Nucleic acid molecules and other molecules associated with the methionine synthesis and degradation pathways

ABSTRACT

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, DNA sequences from maize and soybean plants associated with the methionine pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of applications No. 60/067,000 filedNov. 24, 1997, No. 60/066,873 filed Nov. 25, 1997, No. 60/069,472 filedDec. 9, 1997, No. 60/074,201 filed Feb. 10, 1998, No. 60/074,282 filedFeb. 10, 1998, No. 60/074,280 filed Feb. 10, 1998, No. 60/074,281 filedFeb. 10, 1998, No. 60/074,566 filed Feb. 12, 1998, No. 60/074,567 filedFeb. 12, 1998, No. 60/074,565 filed Feb. 12, 1998, No. 60/075,462 filedFeb. 19, 1998, No. 60/074,789 filed Feb. 19, 1998, No. 60/075,459 filedFeb. 19, 1998, No. 60/075,461 filed Feb. 19, 1998, No. 60/075,464 filedFeb. 19, 1998, No. 60/075,460 filed Feb. 19, 1998, No. 60/075,463 filedFeb. 19, 1998, No. 60/077,231 filed Mar. 9, 1998, No. 60/077,229 filedMar. 9, 1998, No. 60/077,230 filed Mar. 9, 1998, No. 60/078,031 filedMar. 16, 1998, No. 60/078,368 filed Mar. 18, 1998, No. 60/080,844 filedApr. 7, 1998, No. 60/083,067 filed Apr. 27, 1998, “Nucleic AcidMolecules and Other Molecules Associated with Plants” docket No.38-21(15348)A filed Apr. 29, 1998, No. 60/083,387 filed Apr. 29, 1998,No. 60/083,388 filed Apr. 29, 1998, No. 60/083,389 filed Apr. 29, 1998,“Nucleic Acid Molecules and Other Molecules Associated with the EthyleneBiosynthetic Pathway” docket No. 04983.0018/38-21(15097)A filed May 8,1998, No. 60/085,245 filed May 13, 1998, No. 60/085,224 filed May 13,1998, No. 60/085,223 filed May 13, 1998, No. 60/085,222 filed May 13,1998, No. 60/086,186 filed May 21, 1998, No. 60/086,339 filed May 21,1998, No. 60/086,187 filed May 21, 1998, No. 60/086,185 filed May 21,1998, No. 60/086,184 filed May 21, 1998, No. 60/086,183 filed May 21,1998, No. 60/086,188 filed May 21, 1998, No. 60/089,524 filed Jun. 16,1998, No. 60/089,810 filed Jun. 18, 1998, No. 60/089,814 filed Jun. 18,1998, “Nucleic acid molecules and other molecules associated with thePlant Sugar and Nitrogen Transporters Pathway” docket No.04983.0043/38-21(15412)A filed Jun. 30, 1998, No. 60/092,036 filed Jul.8, 1998, No. 60/099,667 filed Sep. 9, 1998, No. 60/099,668 filed Sep. 9,1998, No. 60/099,670 filed Sep. 9, 1998, No. 60/099,697 filed Sep. 9,1998, No. 60/100,674 filed Sep. 16, 1998, No. 60/100,673 filed Sep. 16,1998, No. 60/100,672 filed Sep. 16, 1998, No. 60/101,132 filed Sep. 21,1998, No. 60/101,130 filed Sep. 21, 1998, “Nucleic acid molecules andother molecules associated with Plants” docket No. 38-21(15459)A filedSep. 21, 1998, No. 60/101,344 filed Sep. 22, 1998, No. 60/101,347 filedSep. 22, 1998, No. 60/101,343 filed Sep. 22, 1998, No. 60/104,126 filedOct. 13, 1998, No. 60/104,128 filed Oct. 13, 1998, No. 60/104,127 filedOct. 13, 1998, No. 60/104,124 filed Oct. 13, 1998, “Nucleic AcidMolecules and Other Molecules Associated with Plants” docket No.38-21(15445)A filed Nov. 18, 1998 and “Nucleic Acid Molecules and otherMolecules associated with Plants” docket No. 38-21(15592) filed Nov. 18,1998 hereby incorporated by reference herein in their entirety.

FIELD OF THE INVENTION

The present invention is in the field of plant biochemistry. Morespecifically the invention relates to nucleic acid sequences from plantcells, in particular, DNA sequences from maize and soybean plantsassociated with the methionine pathway. The invention encompassesnucleic acid molecules that encode proteins and fragments of proteins.In addition, the invention also encompasses proteins and fragments ofproteins so encoded and antibodies capable of binding these proteins orfragments. The invention also relates to methods of using the nucleicacid molecules, proteins and fragments of proteins and antibodies, forexample for genome mapping, gene identification and analysis, plantbreeding, preparation of constructs for use in plant gene expression andtransgenic plants.

BACKGROUND OF THE INVENTION

I. Methoinine Synthesis Pathway

The amino acid, L-methionine, is synthesized in higher plants via apathway that starts with L-aspartate. This pathway has been studied(Azevedo et al., Phytochemistry 46:395-419 (1997), the entirety of whichis herein incorporated by reference). L-methionine is one of fourso-called aspartate-derived amino acids (along with L-lysine,L-threonine and L-isoleucine) (Miflin et al., In: Nitrogen Assimilationin Plants, Hewitt et al., (eds.), Academic Press, New York, 335 (1997);Bryan, In: The Biochemistry of Plants, Miflin (ed.), Academic Press, NewYork, 403 (1980); Lea et al., In: The Chemistry and Biochemistry ofAmino Acids, Barrett et al., (eds.), London, 5:197 (1985); Bryan, In:The Biochemistry of Plants, Miflin et al., (eds.), Academic Press, SanDiego, 16:161 (1990), all of which are herein incorporated by referencein their entirety).

The methionine-specific part of the aspartate pathway includes thefollowing enzymes: aspartate kinase (EC 2.7.2.4), aspartate-semialdehydedehydrogenase (EC 1.2.1.11), homoserine dehydrogenase (EC 1.1.1.3),homoserine kinase (EC 2.7.1.39), cystathionine γ-synthase (EC 4.2.99.9),cystathionine β-lyase (EC 4.4.1.8) and methionine synthase (EC2.1.1.14).

Aspartate kinase catalyzes the first reaction of the pathway in whichaspartate is converted to β-aspartyl phosphate. This enzyme has beenisolated and characterized from plant sources including maize, barley,carrot, pea and soybean. These studies have revealed that there aremultiple isoenzymes of aspartate kinase and the isoenzymes differ withrespect to both feedback inhibition sensitivity and expression profile(tissue and developmental stage). Feedback inhibition is mediated bylysine and threonine. Transgenic plants which express an unregulatedaspartate kinase have demonstrated increased flux through the aspartatepathway. Pathway regulation is reported to be exerted, at least in part,via control of this enzyme's activity.

Aspartate semialdehyde dehydrogenase catalyses the second pathwayreaction and converts β-aspartyl phosphate to aspartate semialdehyde viaan NADPH-dependent reaction. Gengenbach et al., Crop Science 18:472-476(1978), the entirety of which is herein incorporated by reference,report the isolation of aspartate semialdehyde dehydrogenase from maizesuspension culture cells. These suspension cultures did not exhibitfeedback inhibition of the enzyme in the presence of aspartate-derivedamino acids, with the exception of methionine, for which some feedbacksensitivity was observed. Aspartate semialdehyde dehydrogenase enzymeactivity has been reported in maize shoot, maize root and maize kernel(Gengenbach et al., Crop Science 18:472-476 (1978)).

Homoserine dehydrogenase catalyzes the next step of the pathway in whichhomoserine is generated from aspartate semialdehyde in a reactionrequiring NADH or NADPH. Homoserine dehydrogenase enzyme has beenstudied in higher plants and multiple isoenzyme forms have been reported(Bryan et al., Biochemistry and Biophysics Research Communications41:1211-1217 (1970); Gengenbach et al., Crop Science 18:472-476 (1978);Dotson et al., Plant Physiology 91:1602-1608 (1989); Dotson et al.,Plant Physiology 93:98-104 (1989); Azevedo et al., Phytochemistry31:3725-3730 (1992); Azevedo et al., Phytochemistry 31:3731-3734 (1992);Brennecke et al., Phytochemistry 41:707 (1996); Aarnes, Plant ScienceLetters 9:137-145 (1977); Bright et al., Biochemical Genetics200:229-243 (1982); Aruda et al., Plant Physiology 76:442-446 (1984);Lea et al., In: Barley: Genetics, Molecular Biology and BiotechnologyShewrey (ed.), CAB International, Oxford 181 (1992); Davies et al.,Plant Science Letters 9:323-332 (1977); Davies et al., Plant Physiology62:536-541 (1978); Matthews et al., Zeitschrift für Naturforschung,Section Bioscience 34:1177-1185 (1979); Relton et al., Biochimica etBiophysica Acta 953:48-60 (1988); Aarnes et al., Phytochemistry13:2717-2724 (1974); Lea et al., FEBS Letters 98:165-168 (1979);Matthews et al., Canadian Journal of Botany 57:299-304 (1979), all ofwhich references are incorporated herein in their entirety). Theisoenzymes have been found to differ with respect to sensitivity tothreonine-mediated feedback inhibition, with both sensitive andinsensitive forms being isolated from maize suspension cultures andseedlings (Miflin et al., In: Nitrogen Assimilation of Plants, Hewitt etal., (eds.), Academic Press, New York, 335 (1997); Bryan, In: TheBiochemistry of Plants, Miflin (ed.), Academic Press, New York, 5:403(1980)).

There is evidence that plants also possess a bifunctional enzyme withboth aspartate kinase and homoserine dehydrogenase activities (Lea etal., In: The Chemistry and Biochemistry of Amino Acids, Barrett et al.(eds), London, 5:197 (1985), the entirety of which is hereinincorporated by reference; Bryan, In: The Biochemistry of Plants, Miflin(ed.), Academic Press, New York, 5:161 (1990), the entirety of which isherein incorporated by reference). Clones of these bifunctional enzymeshave been isolated from Arabidopsis thaliana (Giovanelli et al., In: TheBiochemistry of Plants, Miflin (ed.), Academic Press, New York 453(1990), the entirety of which is herein incorporated by reference)carrot (Giovanelli et al., Plant Physiology 90:1584-1599 (1989), theentirety of which is herein incorporated by reference), maize (Singh etal., Amino Acids 7:165-168 (1994), the entirety of which is hereinincorporated by reference) and soybean (Matthews et al., In:Biosynthesis and Molecular Regulation of Amino Acids in Plants, p 294,Singh et al. (eds.), American Society of Plant Physiologists, Rockville,Md. (1992), the entirety of which is herein incorporated by reference).

The next reported enzymatic step leading to methionine biosynthesis inhigher plants is the final common reaction shared by other amino acidend products (threonine and isoleucine). The reaction is catalyzed byhomoserine kinase and it generates O-phosphohomoserine from homoserine,with ATP serving as the phosphate donor. Exceptions are Pisum sativumand Lathyrus sitivus which synthesize O-acetylhomoserine andO-oxalylhomoserine, respectively (Thomas and Surdin-Kerjan, Microbiol.Mol. Biol. Rev. 61:503-532 (1997), the entirety of which is hereinincorporated by reference). Enteric bacteria use O-succinylhomoserine,while several gram-positive bacteria, yeasts and fungi useO-acetylhomoserine (formed using homoserine O-acetyltransferase (EC2.3.1.31) (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev.61:503-532 (1997)). Homoserine kinase has been reported from multiplehigher plant sources (Galili, The Plant Cell 7:899-906 (1995), theentirety of which is herein incorporated by reference; Rees et al.,Biochemical Journal 309:999-1107 (1995), the entirety of which is hereinincorporated by reference; Bryan et al., Biochemistry and BiophysicsResearch Communications 41:1211-1217 (1970), the entirety of which isherein incorporated by reference; Gengenbach et al., Crop Science18:472-476 (1978), Dotson et al., Plant Physiology 91:1602-1608 (1989),the entirety of which is herein incorporated by reference; Dotson etal., Plant Physiology 93:98-104 (1989), the entirety of which is hereinincorporated by reference). Homoserine kinase isolated from barley andwheat has not been reported to exhibit aspartate-derived amino acidfeedback inhibition (Gengenbach et al., Crop Science 18:472-476 (1978);Dotson et al., Plant Physiology 93:98-104 (1989)). It has been reportedthat homoserine kinase exhibits feedback regulation in the dicots, pea(Rees et al., Biochemical Journal 309:999-1007 (1995), the entirety ofwhich is herein incorporated by reference) and radish (Bryan et al.,Biochemistry and Biophysics Research Communications 41:1211-1217(1970)). Bacterial and yeast homologues have been reported (Azevedo etal., Phytochemistry 31:3725-3730 (1992); Azevedo et al., Phytochemistry31:3731-3734 (1992); Brennecke et al., Phytochemistry 41:707 (1996);Aarnes, Plant Science Letters 9:137-145 (1977)).

Sulfur, in yeast, is incorporated into O-acetylhomoserine resulting inhomocysteine. This reaction is catalyzed by the O-acetylhomoserinesulfhydrylase (EC 4.2.99.10) (also known as O-acethomoserine(thiol)-lyase). O-acetylhomoserine sulfhydrylase has been reported to bea homotetramer with a molecular weight of 200,000. O-acetylhomoserinesulfhydrylase has also been reported to bind four molecules of pyridoxalphosphate (Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev.61:503-532 (1997)).

In higher plants, the sulfur atom from cysteine and the carbon backbonederived from aspartate used to synthesize methionine are reported to becatalyzed by pyridoxal 5′-phosphate (PLP) dependent enzymes (Ravanel etal., Proc. Natl. Acad. Sci. (U.S.A.) 95:7805-7812 (1998), the entiretyof which is herein incorporated by reference). The amino acidcomposition of the O-acetylhomoserine sulfhydrylase has also beenreported to share sequence similarities to the E. coli cystathionineγ-synthase and cystathionine β-lyase and cystathionine γ-lyase fromSaccharomyces cervisiae and rats. All of these enzymes thus appear tobelong to one protein family, whose members have evolved from anancestral pyridoxal phosphate enzyme (Thomas and Surdin-Kerjan,Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

In yeast, the synthesis of cysteine from homocysteine has been reportedto require two successive steps, β addition and γ elimination.Cystathionine β-synthase (EC 4.2.1.22) has been reported to catalyze thefirst reaction where homocysteine and serine yield cystathionine. In S.cervisiae, cystathionine β-synthase is encoded by STR4. STR4 encodes apolypeptide of 506 residues which shows extensive sequence similarity toits functional analog in rats. The rat analog has been reported tocontain an additional amino-terminal extension of 60 residues. Moreover,the two enzymes have been reported to be closely related to the cysteinesynthase from enteric bacteria and plants (Thomas and Surdin-Kerjan,Microbiol. Mol. Biol. Rev. 61:503-532 (1997)).

Cystathionine γ-lyase (EC 4.4.1.1) catalyzes the γ cleavage ofcystationine in yeast, the second reported step of the biosynthesis ofcysteine from homocysteine. Cystathionine γ-lyase has been reported tohave a molecular weight of about 194,000 kd. In S. cerviseae,cystathionine γ-lyase is encoded by STR1. A mutation in the S. cerviseaecystathionine γ-lyase gene leads to a nutritional requirement forcysteine or glutathione. The yeast cystathionine γ-lyase belongs to aprotein family which includes a functional analog in rats, a Met25p fromyeast and cystathionin β-lyase and cystathionin γ-synthase from E. coli(Thomas and Surdin-Kerjan, Microbiol. Mol. Biol. Rev. 61:503-532(1997)).

Cystathionine γ-synthase (also known as O-succinylhomoserine(thio)-lyase, E.C. 4.2.99.9) catalyzes the first reported reaction whichis unique to methionine biosynthesis, thereby committing aspartatepathway flux toward this amino acid. In this reaction,O-phosphohomoserine and cysteine serve as substrates for the productionof cystathionine. Cystathionine γ-synthase has not been reported to beregulated by aspartate-derived amino acids feedback inhibition (Brightet al., Biochemical Genetics 20:229-243 (1982); Arruda et al., PlantPhysiology 76:442-446 (1984)). Cystathionine γ-synthase has however,been reported to be sensitive to product inhibition by orthophosphate(Lea et al., Barley: Genetics, Molecular Biology and Biotechnology,Shewrey (ed.), CAB International, Oxford, 181 (1992); Davies et al.,Plant Science Letters 9:323-332 (1977)). Cloned cystathionine γ-synthasehave been reported from Arabidopsis thaliana (Davies et al., PlantPhysiology 62:536-541 (1978)). It has been reported that methioninelevels are modulated via regulation of cystathionine-synthase (Matthewset al., Zeitschrift für Naturforschung, Section Bioscience 34:1177-1185(1979-2724 (1974); Lea et al., FEBS Letters 98:165 (1979), all of whichreferences are incorporated herein in their entirety).

Cystathionine β-lyase catalyzes the next reaction in the biosynthesis ofmethionine. This reaction generates homocysteine, pyruvate and ammoniafrom the enzymatic decomposition of cystathionine. Evidence forisoenzymes which differ with respect to cellular localization have beenreported for barley (Matthews et al., Canadian Journal of Botany57:299-304 (1979)) and spinach (Rognes et al., Nature 287:357-359(1980), the entirety of which is herein incorporated by reference).

De novo synthesis of methionine from homocysteine uses a methyl groupwhich originates from single-carbon metabolism. In this metabolism,derivatives of tetrahydrofolate transfer one-carbon groups at theoxidation levels of methanol, formaldehyde and formate to acceptormolecules. Single-carbon derivatives of tetrahydrofolate are requiredfor the biosynthesis of methionine, purine nucleotides and thymidylateas well as for the synthesis of N-formylmethionine in the mitochondrion.S. cerevisiae possesses two complete sets of folate interconversionenzymes, one located in the cytosol (methionyl-tRNA synthetase, EC6.1.1.10) and the other located in the mitochondrion (methionyl t-RNAsynthetase, EC 6.1.1.10) (Thomas and Surdin-Kerjan, Microbiol. Mol.Biol. Rev. 61:503-532 (1997)) and in plants including the chloroplast(Menand et al., Proc. Natl. Acad. Sci. (U.S.A.), 95:11014-11019 (1998),the entirety of which is herein incorporated by reference).

Methionine synthase generates methionine from homocysteine by amethylation reaction and thus represents the final step of themethionine biosynthetic pathway. Methionine synthase is also sometimesreferred to as5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferase.N-methyltetrahydrofolate serves as the methyl donor in this reaction,which occurs in the absence of cobalamin (Giovanelli et al., PlantPhysiology 90:1577-1583 (1989), the entirety of which is hereinincorporated by reference; Green et al., Crop Science 14:827-830 (1974),the entirety of which is herein incorporated by reference).

II. Methoinine Degradation Pathway

Plants contain a pathway for the degradation of L-methionine. Thisdegradation pathway includes the following enzymes: methionineadenosyltransferase (EC 2.5.1.6), methionine S-methyltransferase (EC2.1.1.12), adenosylmethionine hydrolase (EC 3.3.1.2), homocysteineS-methyltransferase (EC 2.1.1.10) and S-adenosyl-methioninedecarboxylase (EC 4.1.1.50).

The reported first step in the catabolism of methionine is theATP-dependent conversion to S-adenosylmethionine (AdoMet), which iscatalyzed by the enzyme methionine adenosyltransferase, also known asS-adenosylmethionine synthetase. Methionine adenosyltransferase enzymehas been characterized from several plant sources (Aarnes, Plant ScienceLetters 10:381 (1977), the entirety of which is herein incorporated byreference; Mathur et al., Biochimia and Biophysica Acta 1078:161-170(1991), the entirety of which is herein incorporated by reference; Kimet al., Journal of Biochemical and Molecular Biology 28:100 (1995), theentirety of which is herein incorporated by reference) and nucleic acidmolecules (genomic and cDNA) have also been obtained from a variety ofsources (Izhaki et al., Plant Physiology 108:841-842 (1995), theentirety of which is herein incorporated by reference; Espartero et al.,Molecular Biology Plant 25:217-237 (1994), the entirety of which isherein incorporated by reference). Regulation of methionineadenosyltransferase activity has been observed for the enzyme fromGlycine max (soybean). In Glycine max, methionine adenosyltransferasewas reportedly inhibited by S-adenosylmethionine (Kim et al., Journal ofBiochemical and Molecular Biology 28:100 (1995). Studies have alsoreported that the levels of methionine adenosyltransferase appear tofluctuate in response to hormonal or environmental conditions such asgibberellic acid (Mathur et al., Biochimica and Biophysica Acta1162:289-290 (1993), the entirety of which is herein incorporated byreference; Mathur et al., Biochimica and Biophysica Acta 1137:338-348(1992), the entirety of which is herein incorporated by reference), saltstress (Espartero et al., Molecular Biology Plant 25:217-227 (1994) theentirety of which is herein incorporated by reference) and wounding (Kimet al., Plant Cell Reports 13:340 (1994), the entirety of which isherein incorporated by reference). It has also been reported thatmethionine adenosyltransferase may play a role in the lignificationprocess (Peleman et al., Plant Cell 1:81 (1989), the entirety of whichis herein incorporated by reference).

AdoMet is further catabolized by several enzymes and has been reportedto serve a variety of metabolic functions including that of a methyldonor (Cossins, The Biochemistry of Plants 11:317 Devis (ed.), AcademicPress, San Diego (1987), the entirety of which is herein incorporated byreference) that of a precursor for polyamine biosynthesis (Tiburico etal., The Biochemistry of Plants 16:283 (1990), the entirety of which isherein incorporated by reference) and that of a precursor for ethylenebiosynthesis (Kende, Plant Physiology 91:1-4 (1989), the entirety ofwhich is herein incorporated by reference; Flurh et al., Critical Reviewof Plant Science 15:479 (1996), the entirety of which is hereinincorporated by reference). In each case, enzymes are present toregenerate methionine from the sulfur-containing backbone resulting inno net loss of methionine.

An enzyme involved in AdoMet catabolism is adenosylmethionine hydrolase(EC 3.3.1.2) which converts AdoMet to methylthioadenosine andL-homoserine. L-homoserine is further metabolized during thebiosynthesis of polyamines and ethylene and methylthioadenosine isrecycled to methionine. In yeast, a form of adenosylmethionine hydrolase(EC 3.1.1.1) has been reported(http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query (1998)).

Another enzyme for which AdoMet is a substrate for is homocysteineS-methyltransferase. Homocysteine S-methyltransferase catalyzes thecombination of AdoMet, with L-homocysteine to produce bothS-adenosyl-L-homocysteine and L-methionine. Another enzyme has beendescribed which generates S-adenosyl-L-homocysteine from AdoMet. Thisenzyme is called methionine S-methyltransferase and it catalyzes thereaction in which S-adenosyl-L-homocysteine reacts with L-methionine togenerate S-adenosyl-L-homocysteine and S-methyl-L-methionine. AdoMet canalso be decarboxylated by adenosyl methionine decarboxylase, whichgenerates (5-deoxy-5-adenosyl)(3-aminopropyl) methylsulfonium salt.

III. Expressed Sequence Tag Nucleic Acid Molecules

Expressed sequence tags, or ESTs are randomly sequenced members of acDNA library (or complementary DNA) (McCombie et al., Nature Genetics1:124-130 (1992); Kurata et al., Nature Genetics 8:365-372 (1994); Okuboet al., Nature Genetics 2:173-179 (1992), all of which references areincorporated herein in their entirety). The randomly selected clonescomprise insets that can represent a copy of up to the full length of amRNA transcript.

Using conventional methodologies, cDNA libraries can be constructed fromthe mRNA (messenger RNA) of a given tissue or organism using poly dTprimers and reverse transcriptase (Efstratiadis et al., Cell 7:279-3680(1976), the entirety of which is herein incorporated by reference;Higuchi et al., Proc. Natl. Acad. Sci. (U.S.A.) 73:3146-3150 (1976), theentirety of which is herein incorporated by reference; Maniatis et al.,Cell 8:163-182 (1976) the entirety of which is herein incorporated byreference; Land et al., Nucleic Acids Res. 9:2251-2266 (1981), theentirety of which is herein incorporated by reference; Okayama et al.,Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is hereinincorporated by reference; Gubler et al., Gene 25:263-269 (1983), theentirety of which is herein incorporated by reference).

Several methods may be employed to obtain full-length cDNA constructs.For example, terminal transferase can be used to add homopolymeric tailsof dC residues to the free 3′ hydroxyl groups (Land et al., NucleicAcids Res. 9:2251-2266 (1981), the entirety of which is hereinincorporated by reference). This tail can then be hybridized by a polydG oligo which can act as a primer for the synthesis of full lengthsecond strand cDNA. Okayama and Berg, Mol. Cell. Biol. 2:161-170 (1982),the entirety of which is herein incorporated by reference, report amethod for obtaining full length cDNA constructs. This method has beensimplified by using synthetic primer-adapters that have bothhomopolymeric tails for priming the synthesis of the first and secondstrands and restriction sites for cloning into plasmids (Coleclough etal., Gene 34:305-314 (1985), the entirety of which is hereinincorporated by reference) and bacteriophage vectors (Krawinkel et al.,Nucleic Acids Res. 14:1913 (1986), the entirety of which is hereinincorporated by reference; Han et al., Nucleic Acids Res. 15:6304(1987), the entirety of which is herein incorporated by reference).

These strategies have been coupled with additional strategies forisolating rare mRNA populations. For example, a typical mammalian cellcontains between 10,000 and 30,000 different mRNA sequences (Davidson,Gene Activity in Early Development, 2nd ed., Academic Press, New York(1976), the entirety of which is herein incorporated by reference). Thenumber of clones required to achieve a given probability that alow-abundance mRNA will be present in a cDNA library isN=(ln(1−P))/(ln(1−1/n)) where N is the number of clones required, P isthe probability desired and 1/n is the fractional proportion of thetotal mRNA that is represented by a single rare mRNA (Sambrook et al.,Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring HarborLaboratory Press (1989), the entirety of which is herein incorporated byreference).

A method to enrich preparations of mRNA for sequences of interest is tofractionate by size. One such method is to fractionate byelectrophoresis through an agarose gel (Pennica et al., Nature301:214-221 (1983), the entirety of which is herein incorporated byreference). Another such method employs sucrose gradient centrifugationin the presence of an agent, such as methylmercuric hydroxide, thatdenatures secondary structure in RNA (Schweinfest et al., Proc. Natl.Acad. Sci. (U.S.A.) 79:4997-5000 (1982), the entirety of which is hereinincorporated by reference).

A frequently adopted method is to construct equalized or normalized cDNAlibraries (Ko, Nucleic Acids Res. 18:5705-5711 (1990), the entirety ofwhich is herein incorporated by reference; Patanjali et al., Proc. Natl.Acad. Sci. (U.S.A.) 88:1943-1947 (1991), the entirety of which is hereinincorporated by reference). Typically, the cDNA population is normalizedby subtractive hybridization (Schmid et al., J. Neurochem. 48:307-312(1987), the entirety of which is herein incorporated by reference;Fargnoli et al., Anal. Biochem. 187:364-373 (1990), the entirety ofwhich is herein incorporated by reference; Travis et al., Proc. Natl.Acad. Sci. (U.S.A.) 85:1696-1700 (1988), the entirety of which is hereinincorporated by reference; Kato, Eur. J. Neurosci. 2:704-711 (1990); andSchweinfest et al., Genet. Anal. Tech. Appl. 7:64-70 (1990), theentirety of which is herein incorporated by reference). Subtractionrepresents another method for reducing the population of certainsequences in the cDNA library (Swaroop et al., Nucleic Acids Res.19:1954 (1991), the entirety of which is herein incorporated byreference).

ESTs can be sequenced by a number of methods. Two basic methods may beused for DNA sequencing, the chain termination method of Sanger et al.,Proc. Natl. Acad. Sci. (U.S.A.) 74:5463-5467 (1977), the entirety ofwhich is herein incorporated by reference and the chemical degradationmethod of Maxam and Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564(1977), the entirety of which is herein incorporated by reference.Automation and advances in technology such as the replacement ofradioisotopes with fluorescence-based sequencing have reduced the effortrequired to sequence DNA (Craxton, Methods 2:20-26 (1991), the entiretyof which is herein incorporated by reference; Ju et al., Proc. Natl.Acad. Sci. (U.S.A.) 92:4347-4351 (1995), the entirety of which is hereinincorporated by reference; Tabor and Richardson, Proc. Natl. Acad. Sci.(U.S.A.) 92:6339-6343 (1995), the entirety of which is hereinincorporated by reference). Automated sequencers are available from, forexample, Pharmacia Biotech, Inc., Piscataway, N.J. (Pharmacia ALF),LI-COR, Inc., Lincoln, Nebr. (LI-COR 4,000) and Millipore, Bedford,Mass. (Millipore BaseStation).

In addition, advances in capillary gel electrophoresis have also reducedthe effort required to sequence DNA and such advances provide a rapidhigh resolution approach for sequencing DNA samples (Swerdlow andGesteland, Nucleic Acids Res. 18:1415-1419 (1990); Smith, Nature349:812-813 (1991); Luckey et al., Methods Enzymol. 218:154-172 (1993);Lu et al., J. Chromatog. A. 680:497-501 (1994); Carson et al., Anal.Chem. 65:3219-3226 (1993); Huang et al., Anal. Chem. 64:2149-2154(1992); Kheterpal et al., Electrophoresis 17:1852-1859 (1996); Quesadaand Zhang, Electrophoresis 17:1841-1851 (1996); Baba, Yakugaku Zasshi117:265-281 (1997), all of which are herein incorporated by reference intheir entirety).

ESTs longer than 150 nucleotides have been found to be useful forsimilarity searches and mapping (Adams et al., Science 252:1651-1656(1991), herein incorporated by reference). ESTs, which can representcopies of up to the full length transcript, may be partially orcompletely sequenced. Between 150-450 nucleotides of sequenceinformation is usually generated as this is the length of sequenceinformation that is routinely and reliably produced using single runsequence data. Typically, only single run sequence data is obtained fromthe cDNA library (Adams et al., Science 252:1651-1656 (1991). Automatedsingle run sequencing typically results in an approximately 2-3% erroror base ambiguity rate (Boguski et al., Nature Genetics 4:332-333(1993), the entirety of which is herein incorporated by reference).

EST databases have been constructed or partially constructed from, forexample, C. elegans (McCombrie et al., Nature Genetics 1: 124-131(1992)), human liver cell line HepG2 (Okubo et al., Nature Genetics2:173-179 (1992)), human brain RNA (Adams et al., Science 252:1651-1656(1991); Adams et al., Nature 355:632-635 (1992)), Arabidopsis, (Newmanet al., Plant Physiol. 106:1241-1255 (1994)); and rice (Kurata et al.,Nature Genetics 8:365-372 (1994)).

IV. Sequence Comparisons

A characteristic feature of a DNA sequence is that it can be comparedwith other DNA sequences. Sequence comparisons can be undertaken bydetermining the similarity of the test or query sequence with sequencesin publicly available or proprietary databases (“similarity analysis”)or by searching for certain motifs (“intrinsic sequence analysis”) (e.g.cis elements) (Coulson, Trends in Biotechnology 12:76-80 (1994), theentirety of which is herein incorporated by reference); Birren et al.,Genome Analysis 1: Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y. 543-559 (1997), the entirety of which is hereinincorporated by reference).

Similarity analysis includes database search and alignment. Examples ofpublic databases include the DNA Database of Japan (DDBJ)(http://www.ddbj.nig.ac.jp/); Genebank(http://www.ncbi.nlm.nih.gov/Web/Search/Index.htlm); and the EuropeanMolecular Biology Laboratory Nucleic Acid Sequence Database (EMBL)(http://www.ebi.ac.uk/ebi_docs/embl_db/embl-db.html). Other appropriatedatabases include dbEST (http://www.ncbi.nlm.nih.gov/dbEST/index.html),SwissProt (http://www.ebi.ac.uk/ebi_docs/swisprot_db/swisshome.html),PIR (http://www-nbrt.georgetown.edu/pir/) and The Institute for GenomeResearch (http://www.tigr.org/tdb/tdb.html)

A number of different search algorithms have been developed, one exampleof which are the suite of programs referred to as BLAST programs. Thereare five implementations of BLAST, three designed for nucleotidesequences queries (BLASTN, BLASTX and TBLASTX) and two designed forprotein sequence queries (BLASTP and TBLASTN) (Coulson, Trends inBiotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1, ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559(1997)).

BLASTN takes a nucleotide sequence (the query sequence) and its reversecomplement and searches them against a nucleotide sequence database.BLASTN was designed for speed, not maximum sensitivity and may not finddistantly related coding sequences. BLASTX takes a nucleotide sequence,translates it in three forward reading frames and three reversecomplement reading frames and then compares the six translations againsta protein sequence database. BLASTX is useful for sensitive analysis ofpreliminary (single-pass) sequence data and is tolerant of sequencingerrors (Gish and States, Nature Genetics 3:266-272 (1993), the entiretyof which is herein incorporated by reference). BLASTN and BLASTX may beused in concert for analyzing EST data (Coulson, Trends in Biotechnology12:76-80 (1994); Birren et al., Genome Analysis 1:543-559 (1997)).

Given a coding nucleotide sequence and the protein it encodes, it isoften preferable to use the protein as the query sequence to search adatabase because of the greatly increased sensitivity to detect moresubtle relationships. This is due to the larger alphabet of proteins (20amino acids) compared with the alphabet of nucleic acid sequences (4bases), where it is far easier to obtain a match by chance. In addition,with nucleotide alignments, only a match (positive score) or a mismatch(negative score) is obtained, but with proteins, the presence ofconservative amino acid substitutions can be taken into account. Here, amismatch may yield a positive score if the non-identical residue hasphysical/chemical properties similar to the one it replaced. Variousscoring matrices are used to supply the substitution scores of allpossible amino acid pairs. A general purpose scoring system is theBLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61 (1993), theentirety of which is herein incorporated by reference), which iscurrently the default choice for BLAST programs. BLOSUM62 is tailoredfor alignments of moderately diverged sequences and thus may not yieldthe best results under all conditions. Altschul, J. Mol. Biol.36:290-300 (1993), the entirety of which is herein incorporated byreference, describes a combination of three matrices to cover allcontingencies. This may improve sensitivity, but at the expense ofslower searches. In practice, a single BLOSUM62 matrix is often used butothers (PAM40 and PAM250) may be attempted when additional analysis isnecessary. Low PAM matrices are directed at detecting very strong butlocalized sequence similarities, whereas high PAM matrices are directedat detecting long but weak alignments between very distantly relatedsequences.

Homologues in other organisms are available that can be used forcomparative sequence analysis. Multiple alignments are performed tostudy similarities and differences in a group of related sequences.CLUSTAL W is a multiple sequence alignment package that performsprogressive multiple sequence alignments based on the method of Feng andDoolittle, J. Mol. Evol. 25:351-360 (1987), the entirety of which isherein incorporated by reference. Each pair of sequences is aligned andthe distance between each pair is calculated; from this distance matrix,a guide tree is calculated and all of the sequences are progressivelyaligned based on this tree. A feature of the program is its sensitivityto the effect of gaps on the alignment; gap penalties are varied toencourage the insertion of gaps in probable loop regions instead of inthe middle of structured regions. Users can specify gap penalties,choose between a number of scoring matrices, or supply their own scoringmatrix for both pairwise alignments and multiple alignments. CLUSTAL Wfor UNIX and VMS systems is available at: ftp.ebi.ac.uk. Another programis MACAW (Schuler et al., Proteins Struct. Func. Genet. 9:180-190(1991), the entirety of which is herein incorporated by reference, forwhich both Macintosh and Microsoft Windows versions are available. MACAWuses a graphical interface, provides a choice of several alignmentalgorithms and is available by anonymous ftp at: ncbi.nlm.nih.gov(directory/pub/macaw).

Sequence motifs are derived from multiple alignments and can be used toexamine individual sequences or an entire database for subtle patterns.With motifs, it is sometimes possible to detect distant relationshipsthat may not be demonstrable based on comparisons of primary sequencesalone. Currently, the largest collection of sequence motifs in the worldis PROSITE (Bairoch and Bucher, Nucleic Acid Research 22:3583-3589(1994), the entirety of which is herein incorporated by reference).PROSITE may be accessed via either the ExPASy server on the World WideWeb or anonymous ftp site. Many commercial sequence analysis packagesalso provide search programs that use PROSITE data.

A resource for searching protein motifs is the BLOCKS E-mail serverdeveloped by Henikoff, Trends Biochem Sci. 18:267-268 (1993), theentirety of which is herein incorporated by reference; Henikoff andHenikoff, Nucleic Acid Research 19:6565-6572 (1991), the entirety ofwhich is herein incorporated by reference; Henikoff and Henikoff,Proteins 17:49-61 (1993). BLOCKS searches a protein or nucleotidesequence against a database of protein motifs or “blocks.” Blocks aredefined as short, ungapped multiple alignments that represent highlyconserved protein patterns. The blocks themselves are derived fromentries in PROSITE as well as other sources. Either a protein query or anucleotide query can be submitted to the BLOCKS server; if a nucleotidesequence is submitted, the sequence is translated in all six readingframes and motifs are sought for these conceptual translations. Once thesearch is completed, the server will return a ranked list of significantmatches, along with an alignment of the query sequence to the matchedBLOCKS entries.

Conserved protein domains can be represented by two-dimensionalmatrices, which measure either the frequency or probability of theoccurrences of each amino acid residue and deletions or insertions ineach position of the domain. This type of model, when used to searchagainst protein databases, is sensitive and usually yields more accurateresults than simple motif searches. Two popular implementations of thisapproach are profile searches such as GCG program ProfileSearch andHidden Markov Models (HMMs) (Krough et al., J. Mol. Biol. 235:1501-1531,(1994); Eddy, Current Opinion in Structural Biology 6:361-365, (1996),both of which are herein incorporated by reference in their entirety).In both cases, a large number of common protein domains have beenconverted into profiles, as present in the PROSITE library, or HHMmodels, as in the Pfam protein domain library (Sonnhammer et al.,Proteins 28:405-420 (1997), the entirety of which is herein incorporatedby reference). Pfam contains more than 500 HMM models for enzymes,transcription factors, signal transduction molecules and structuralproteins. Protein databases can be queried with these profiles or HMMmodels, which will identify proteins containing the domain of interest.For example, HMMSW or HMMFS, two programs in a public domain packagecalled HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can beused.

PROSITE and BLOCKS represent collected families of protein motifs. Thus,searching these databases entails submitting a single sequence todetermine whether or not that sequence is similar to the members of anestablished family. Programs working in the opposite direction compare acollection of sequences with individual entries in the proteindatabases. An example of such a program is the Motif Search Tool, orMoST (Tatusov et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:12091-12095(1994), the entirety of which is herein incorporated by reference). Onthe basis of an aligned set of input sequences, a weight matrix iscalculated by using one of four methods (selected by the user). A weightmatrix is simply a representation, position by position of how likely aparticular amino acid will appear. The calculated weight matrix is thenused to search the databases. To increase sensitivity, newly foundsequences are added to the original data set, the weight matrix isrecalculated and the search is performed again. This procedure continuesuntil no new sequences are found.

SUMMARY OF THE INVENTION

The present invention provides a substantially purified nucleic acidmolecule that encodes a maize or soybean enzyme or fragment thereof,wherein said maize or soybean enzyme is selected from the groupconsisting of: (a) methionine adenosyltransferase, (b)S-adenosyl-methionine decarboxylase, (c) aspartate kinase, (d)aspartate-semialdehyde dehydrogenase, (e) cystathionine gamma-synthase,(f) cystathionine beta-lysase, and (g)5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferase.

The present invention also provides a substantially purified nucleicacid molecule that encodes a plant methionine pathway enzyme or fragmentthereof, wherein the nucleic acid molecule is selected from the groupconsisting of a nucleic acid molecule that encodes a maize or a soybeanmethionine adenosyltransferase enzyme or fragment thereof, a nucleicacid molecule that encodes a maize or a soybean S-adenosylmethioninedecarboxylase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean aspartate kinase enzyme or fragmentthereof, a nucleic acid molecule that encodes a maize or a soybeanaspartate-semialdehyde dehydrogenase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybeanO-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleicacid molecule that encodes a maize or a soybean cystathionine β-lyaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean adenosylhomocysteinase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof.

A substantially purified maize or soybean enzyme or fragment thereof,wherein said maize or soybean enzyme is selected from the groupconsisting of (a) methionine adenosyltransferase or fragment thereof;(b) S-adenosyl-methionine decarboxylase or fragment thereof; (c)aspartate kinase or fragment thereof; (d) aspartate-semialdehydedehydrogenase or fragment thereof; (e) cystathionine gamma-synthase orfragment thereof; (f) cystathionine beta-lysase or fragment thereof; and(e)5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferaseor fragment thereof.

The present invention also provides a substantially purified maize orsoybean methionine pathway protein or fragment thereof encoded by afirst nucleic acid molecule which specifically hybridizes to a secondnucleic acid molecule, the second nucleic acid molecule having a nucleicacid sequence selected from the group consisting of a complement of SEQID NO: 1 through SEQ ID NO: 3204.

The present invention also provides a substantially purified maize orsoybean methionine adenosyltransferase enzyme or fragment thereofencoded by a first nucleic acid molecule which specifically hybridizesto a second nucleic acid molecule, the second nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of acomplement of SEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635through SEQ ID NO: 2479.

The present invention also provides a substantially purified maize orsoybean methionine adenosyltransferase enzyme or fragment thereofencoded by a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ IDNO: 2479.

The present invention also provides a substantially purified maize orsoybean S-adenosylmethionine decarboxylase enzyme or fragment thereofencoded by a first nucleic acid molecule which specifically hybridizesto a second nucleic acid molecule, the second nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of acomplement of SEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480through SEQ ID NO: 2623.

The present invention also provides a substantially purified maize orsoybean S-adenosylmethionine decarboxylase enzyme or fragment thereofencoded by a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ IDNO: 2623.

The present invention also provides a substantially purified maize orsoybean aspartate kinase enzyme or fragment thereof encoded by a firstnucleic acid molecule which specifically hybridizes to a second nucleicacid molecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO:2648.

The present invention also provides a substantially purified maize orsoybean aspartate kinase enzyme or fragment thereof encoded by a nucleicacid sequence selected from the group consisting of SEQ ID NO: 858through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ ID NO: 2648.

The present invention also provides a substantially purified maize orsoybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereofencoded by a first nucleic acid molecule which specifically hybridizesto a second nucleic acid molecule, the second nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of acomplement of SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649through SEQ ID NO: 2654.

The present invention also provides a substantially purified maize orsoybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereofencoded by a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ IDNO: 2654.

The present invention also provides a substantially purified maize orsoybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereofencoded by a first nucleic acid molecule which specifically hybridizesto a second nucleic acid molecule, the second nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of acomplement of SEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655through SEQ ID NO: 2660.

The present invention also provides a substantially purified maize orsoybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereofencoded by a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ IDNO: 2660.

The present invention also provides a substantially purified maize orsoybean cystathionine β-lyase enzyme or fragment thereof encoded by afirst nucleic acid molecule which specifically hybridizes to a secondnucleic acid molecule, the second nucleic acid molecule having a nucleicacid sequence selected from the group consisting of a complement of SEQID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO:2665.

The present invention also provides a substantially purified maize orsoybean cystathionine β-lyase enzyme or fragment thereof encoded by anucleic acid sequence selected from the group consisting of SEQ ID NO:954 through SEQ ID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665.

The present invention also provides a substantially purified maize orsoybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 964 through SEQID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified maize orsoybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof encoded by a nucleic acid sequence selectedfrom the group consisting of SEQ ID NO: 964 through SEQ ID NO: 1353 andSEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified maize oradenosylhomocysteinase enzyme or fragment thereof encoded by a firstnucleic acid molecule which specifically hybridizes to a second nucleicacid molecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 1354 through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO:3199.

The present invention also provides a substantially purified maize oradenosylhomocysteinase enzyme or fragment thereof encoded by a nucleicacid sequence selected from the group consisting of SEQ ID NO: 1354through SEQ ID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199.

The present invention also provides a substantially purified maize orcystathionine β-synthase enzyme or fragment thereof encoded by a firstnucleic acid molecule which specifically hybridizes to a second nucleicacid molecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 1631 through SEQ ID NO: 1632.

The present invention also provides a substantially purified maize orcystathionine β-synthase enzyme or fragment thereof encoded by a nucleicacid sequence selected from the group consisting of SEQ ID NO: 1631through SEQ ID NO: 1632.

The present invention also provides a substantially purified maize orcystathionine γ-lyase enzyme or fragment thereof encoded by a firstnucleic acid molecule which specifically hybridizes to a second nucleicacid molecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 1633 through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO:3204.

The present invention also provides a substantially purified maize orcystathionine γ-lyase enzyme or fragment thereof encoded by a nucleicacid sequence selected from the group consisting of SEQ ID NO: 1633through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204.

The present invention also provides a substantially purified maize orO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by afirst nucleic acid molecule which specifically hybridizes to a secondnucleic acid molecule, the second nucleic acid molecule having a nucleicacid sequence selected from the group consisting of a complement of SEQID NO: 3200 through SEQ ID NO: 3202.

The present invention also provides a substantially purified maize orO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof encoded by anucleic acid sequence selected from the group consisting of SEQ ID NO:3200 through SEQ ID NO: 3202.

The present invention also provides a purified antibody or fragmentthereof which is capable of specifically binding to a specific maize orsoybean enzyme or fragment thereof, wherein said maize or soybean enzymeor fragment thereof is encoded by a nucleic acid molecule comprising anucleic acid sequence selected from the group consisting of consistingof SEQ ID NO: 1 through SEQ ID NO: 3204.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean methionineadenosyltransferase enzyme or fragment thereof encoded by a firstnucleic acid molecule which specifically hybridizes to a second nucleicacid molecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 1 through SEQ ID NO: 429 and SEQ. ID NO: 1635 through SEQ ID NO:2479 or a substantially purified maize or soybean methionineadenosyltransferase enzyme or fragment thereof encoded by a nucleic acidsequence selected from the group consisting of SEQ ID NO: 1 through SEQID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean S-adenosylmethioninedecarboxylase enzyme or fragment thereof encoded by a first nucleic acidmolecule which specifically hybridizes to a second nucleic acidmolecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO:2623 or a substantially purified maize or soybean S-adenosylmethioninedecarboxylase enzyme or fragment thereof encoded by a nucleic acidsequence selected from the group consisting of SEQ ID NO: 430 throughSEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean aspartate kinase enzyme orfragment thereof encoded by a first nucleic acid molecule whichspecifically hybridizes to a second nucleic acid molecule, the secondnucleic acid molecule having a nucleic acid sequence selected from thegroup consisting of a complement of SEQ ID NO: 858 through SEQ ID NO:900 and SEQ ID NO: 2624 through SEQ ID NO: 2648 or a substantiallypurified maize or soybean aspartate kinase enzyme or fragment thereofencoded by a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ IDNO: 2648.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean aspartate-semialdehydedehydrogenase enzyme or fragment thereof encoded by a first nucleic acidmolecule which specifically hybridizes to a second nucleic acidmolecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO:2654 or a substantially purified maize or soybean enzymeaspartate-semialdehyde dehydrogenase or fragment thereof encoded by anucleic acid sequence selected from the group consisting of SEQ ID NO:901 through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean O-succinylhomoserine(thiol)-lyase enzyme or fragment thereof encoded by a first nucleic acidmolecule which specifically hybridizes to a second nucleic acidmolecule, the second nucleic acid molecule having a nucleic acidsequence selected from the group consisting of a complement of SEQ IDNO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO:2660 or a substantially purified maize or soybean O-succinylhomoserine(thiol)-lyase enzyme or fragment thereof encoded by a nucleic acidsequence selected from the group consisting of SEQ ID NO: 905 throughSEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO: 2660.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean cystathionine β-lyaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 954 through SEQID NO: 963 and SEQ ID NO: 2661 through SEQ ID NO: 2665 or asubstantially purified maize or soybean cystathionine β-lyase enzyme orfragment thereof encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO:2661 through SEQ ID NO: 2665.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 964 through SEQID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992 or asubstantially purified maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof encoded by a nucleic acid sequence selectedfrom the group consisting of SEQ ID NO: 964 through SEQ ID NO: 1353 andSEQ ID NO: 2666 through SEQ ID NO: 2992.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean adenosylhomocyteinaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 1354 through SEQID NO: 1630 and SEQ ID NO: 2993 through SEQ ID NO: 3199 or asubstantially purified maize or soybean adenosylhomocyteinase enzyme orfragment thereof encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 1354 through SEQ ID NO: 1630 and SEQ IDNO: 2993 through SEQ ID NO: 3199.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean cystathionine β-synthaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 1631 through SEQID NO: 1632 or a substantially purified maize or soybean cystathionineβ-synthase enzyme or fragment thereof encoded by a nucleic acid sequenceselected from the group consisting of SEQ ID NO: 1631 through SEQ ID NO:1632.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean cystathionine γ-lyaseenzyme or fragment thereof encoded by a first nucleic acid moleculewhich specifically hybridizes to a second nucleic acid molecule, thesecond nucleic acid molecule having a nucleic acid sequence selectedfrom the group consisting of a complement of SEQ ID NO: 1633 through SEQID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 or asubstantially purified maize or soybean cystathionine γ-lyase enzyme orfragment thereof encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 1633 through SEQ ID NO: 1634 and SEQ IDNO: 3203 through SEQ ID NO: 3204.

The present invention also provides a substantially purified antibody orfragment thereof, the antibody or fragment thereof capable ofspecifically binding to a maize or a soybean O-acetylhomoserine enzymeor fragment thereof encoded by a first nucleic acid molecule whichspecifically hybridizes to a second nucleic acid molecule, the secondnucleic acid molecule having a nucleic acid sequence selected from thegroup consisting of a complement of SEQ ID NO: 3200 through SEQ ID NO:3202 or a substantially purified maize or soybean O-acetylhomoserineenzyme or fragment thereof encoded by a nucleic acid sequence selectedfrom the group consisting of SEQ ID NO: 3200 through SEQ ID NO: 3202.

The present invention also provides a transformed plant having a nucleicacid molecule which comprises: (A) an exogenous promoter region whichfunctions in a plant cell to cause the production of a mRNA molecule;(B) a structural nucleic acid molecule comprising a nucleic acidsequence selected from the group consisting of (a) a nucleic acidsequence which encodes for methionine adenosyltransferase or fragmentthereof, (b) a nucleic acid sequence which encodes forS-adenosyl-methionine decarboxylase or fragment thereof, (c) a nucleicacid sequence which encodes for aspartate kinase or fragment thereof;(d) a nucleic acid sequence which encodes for aspartate-semialdehydedehydrogenase or fragment thereof; (e) a nucleic acid sequence whichencodes for cystathionine gamma-synthase or a fragment thereof; (f) anucleic acid sequence which encodes for cystathionine beta-lysase or afragment thereof; (g) a nucleic acid sequence which encodes for5-methyltetrahydropteroyl-triglutamate-homocysteine-S-methyltransferaseor a fragment thereof; and (h) a nucleic acid sequence which iscomplementary to any of the nucleic acid sequences of (a) through (g);and (C) a 3′ non-translated sequence that functions in said plant cellto cause termination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of said mRNA molecule.

The present invention also provides a transformed plant having a nucleicacid molecule which comprises: (A) an exogenous promoter region whichfunctions in a plant cell to cause the production of a mRNA molecule;which is linked to (B) a structural nucleic acid molecule, wherein thestructural nucleic acid molecule encodes a plant methionine pathwayenzyme or fragment thereof, the structural nucleic acid moleculecomprising a nucleic acid sequence selected from the group consisting ofSEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof; which islinked to (C) a 3′ non-translated sequence that functions in the plantcell to cause termination of transcription and addition ofpolyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleicacid molecule which comprises: (A) an exogenous promoter region whichfunctions in a plant cell to cause the production of a mRNA molecule;which is linked to (B) a structural nucleic acid molecule, wherein thestructural nucleic acid molecule is selected from the group consistingof a nucleic acid molecule that encodes a maize or a soybean methionineadenosyltransferase enzyme or fragment thereof, a nucleic acid moleculethat encodes a maize or a soybean S-adenosylmethionine decarboxylaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor soybean aspartate kinase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean aspartate-semialdehydedehydrogenase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzymeor fragment thereof, a nucleic acid molecule that encodes a maize or asoybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean adenosylhomocysteinase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; which islinked to (C) a 3′ non-translated sequence that functions in the plantcell to cause termination of transcription and addition ofpolyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleicacid molecule which comprises: (A) an exogenous promoter region whichfunctions in a plant cell to cause the production of a mRNA molecule;which is linked to (B) a transcribed nucleic acid molecule with atranscribed strand and a non-transcribed strand, wherein the transcribedstrand is complementary to a nucleic acid molecule comprising a nucleicacid sequence selected from the group consisting of SEQ ID NO: 1 throughSEQ ID NO: 3204 or fragment thereof; which is linked to (C) a 3′non-translated sequence that functions in plant cells to causetermination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleicacid molecule which comprises: (A) an exogenous promoter region whichfunctions in a plant cell to cause the production of a mRNA molecule;which is linked to: (B) a transcribed nucleic acid molecule with atranscribed strand and a non-transcribed strand, wherein a transcribedmRNA of the transcribed strand is complementary to an endogenous mRNAmolecule having a nucleic acid sequence selected from the groupconsisting of an endogenous mRNA molecule that encodes a maize or asoybean methionine adenosyltransferase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybeanS-adenosylmethionine decarboxylase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybean aspartatekinase enzyme or fragment thereof, an endogenous mRNA molecule thatencodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzymeor fragment thereof, an endogenous mRNA molecule that encodes a maize ora soybean O-succinylhomoserine (thiol)-lyase enzyme or fragment thereof,an endogenous mRNA molecule that encodes a maize or a soybeancystathionine β-lyase enzyme or fragment thereof, an endogenous mRNAmolecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferaseenzyme or fragment thereof, an endogenous mRNA molecule that encodes amaize or a soybean adenosylhomocysteinase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, an endogenous mRNA molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and an endogenous mRNA molecule that encodes a maize or asoybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof;which is linked to (C) a 3′ non-translated sequence that functions inthe plant cell to cause termination of transcription and addition ofpolyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a method for determining a level orpattern in a plant cell of an enzyme in a plant metabolic pathwaycomprising: (A) incubating, under conditions permitting nucleic acidhybridization, a marker nucleic acid molecule, said marker nucleic acidmolecule selected from the group of marker nucleic acid molecules whichspecifically hybridize to a nucleic acid molecule having the nucleicacid sequence of SEQ ID NO: 1 through SEQ ID NO: 3204 or complimentsthereof, with a complementary nucleic acid molecule obtained from saidplant cell or plant tissue, wherein nucleic acid hybridization betweensaid marker nucleic acid molecule and said complementary nucleic acidmolecule obtained from said plant cell or plant tissue permits thedetection of an mRNA for said enzyme; (B) permitting hybridizationbetween said marker nucleic acid molecule and said complementary nucleicacid molecule obtained from said plant cell or plant tissue; and (C)detecting the level or pattern of said complementary nucleic acid,wherein the detection of said complementary nucleic acid is predictiveof the level or pattern of said enzyme in said plant metabolic pathway.

The present invention also provides a method for determining a level orpattern of a plant methionine pathway enzyme in a plant cell or planttissue comprising: (A) incubating, under conditions permitting nucleicacid hybridization, a marker nucleic acid molecule, the marker nucleicacid molecule having a nucleic acid sequence selected from the groupconsisting of SEQ ID NO: 1 through SEQ ID NO: 3204 or complement thereofor fragment of either, with a complementary nucleic acid moleculeobtained from the plant cell or plant tissue, wherein nucleic acidhybridization between the marker nucleic acid molecule and thecomplementary nucleic acid molecule obtained from the plant cell orplant tissue permits the detection of the plant methionine pathwayenzyme; (B) permitting hybridization between the marker nucleic acidmolecule and the complementary nucleic acid molecule obtained from theplant cell or plant tissue; and (C) detecting the level or pattern ofthe complementary nucleic acid, wherein the detection of thecomplementary nucleic acid is predictive of the level or pattern of theplant methionine pathway enzyme.

The present invention also provides a method for determining a level orpattern of a plant methionine pathway enzyme in a plant cell or planttissue comprising: (A) incubating, under conditions permitting nucleicacid hybridization, a marker nucleic acid molecule, the marker nucleicacid molecule comprising a nucleic acid molecule that encodes a maize ora soybean methionine adenosyltransferase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean S-adenosylmethionine decarboxylase enzyme or complement thereofor fragment of either, a nucleic acid molecule that encodes a maize or asoybean aspartate kinase enzyme or complement thereof or fragment ofeither, a nucleic acid molecule that encodes a maize or a soybeanaspartate-semialdehyde dehydrogenase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereofor fragment of either, a nucleic acid molecule that encodes a maize or asoybean cystathionine β-lyase enzyme or complement thereof or fragmentof either, a nucleic acid molecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or a soybean adenosylhomocysteinase enzymeor complement thereof or fragment of either, a nucleic acid moleculethat encodes a maize or a soybean cystathionine β-synthase enzyme orcomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean cytsathionine γ-lyase enzyme or complementthereof or fragment of either and a nucleic acid molecule that encodes amaize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complementthereof or fragment of either, with a complementary nucleic acidmolecule obtained from the plant cell or plant tissue, wherein nucleicacid hybridization between the marker nucleic acid molecule and thecomplementary nucleic acid molecule obtained from the plant cell orplant tissue permits the detection of the plant methionine pathwayenzyme; (B) permitting hybridization between the marker nucleic acidmolecule and the complementary nucleic acid molecule obtained from theplant cell or plant tissue; and (C) detecting the level or pattern ofthe complementary nucleic acid, wherein the detection of thecomplementary nucleic acid is predictive of the level or pattern of theplant methionine pathway enzyme.

The present invention also provides a method for determining a level orpattern of a plant methionine pathway enzyme in a plant cell or planttissue under evaluation which comprises assaying the concentration of amolecule, whose concentration is dependent upon the expression of agene, the gene specifically hybridizes to a nucleic acid molecule havinga nucleic acid sequence selected from the group consisting of SEQ ID NO:1 through SEQ ID NO: 3204 or complements thereof, in comparison to theconcentration of that molecule present in a reference plant cell or areference plant tissue with a known level or pattern of the plantmethionine pathway enzyme, wherein the assayed concentration of themolecule is compared to the assayed concentration of the molecule in thereference plant cell or reference plant tissue with the known level orpattern of the plant methionine pathway enzyme.

The present invention also provides a method for determining a level orpattern of a plant methionine pathway enzyme in a plant cell or planttissue under evaluation which comprises assaying the concentration of amolecule, whose concentration is dependent upon the expression of agene, the gene specifically hybridizes to a nucleic acid moleculeselected from the group consisting of a nucleic acid molecule thatencodes a maize or a soybean methionine adenosyltransferase enzyme orcomplement thereof, a nucleic acid molecule that encodes a maize or asoybean S-adenosylmethionine decarboxylase enzyme or complement thereof,a nucleic acid molecule that encodes a maize or a soybean aspartatekinase enzyme or complement thereof, a nucleic acid molecule thatencodes a maize or a soybean aspartate-semialdehyde dehydrogenase enzymeor complement thereof, a nucleic acid molecule that encodes a maize or asoybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof,a nucleic acid molecule that encodes a maize or a soybean cystathionineβ-lyase enzyme or complement thereof, a nucleic acid molecule thatencodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof, a nucleic acid molecule that encodes amaize or a soybean adenosylhomocyteinase enzyme or complement thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or complement thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or complementthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or complement thereof, incomparison to the concentration of that molecule present in a referenceplant cell or a reference plant tissue with a known level or pattern ofthe plant methionine pathway enzyme, wherein the assayed concentrationof the molecule is compared to the assayed concentration of the moleculein the reference plant cell or the reference plant tissue with the knownlevel or pattern of the plant methionine pathway enzyme.

A method of determining a mutation in a plant whose presence ispredictive of a mutation affecting a level or pattern of a proteincomprising the steps: (A) incubating, under conditions permittingnucleic acid hybridization, a marker nucleic acid, said marker nucleicacid selected from the group of marker nucleic acid molecules whichspecifically hybridize to a nucleic acid molecule having a nucleic acidsequence selected from the group of SEQ ID NO: 1 through SEQ ID NO: 3204or complements thereof and a complementary nucleic acid moleculeobtained from said plant, wherein nucleic acid hybridization betweensaid marker nucleic acid molecule and said complementary nucleic acidmolecule obtained from said plant permits the detection of apolymorphism whose presence is predictive of a mutation affecting saidlevel or pattern of said plant methionine pathway enzyme in said plant;(B) permitting hybridization between said marker nucleic acid moleculeand said complementary nucleic acid molecule obtained from said plant;and (C) detecting the presence of said polymorphism, wherein thedetection of said polymorphism is predictive of said mutation.

The present invention also provides a method for determining a mutationin a plant whose presence is predictive of a mutation affecting thelevel or pattern of a plant methionine pathway enzyme comprising thesteps: (A) incubating, under conditions permitting nucleic acidhybridization, a marker nucleic acid molecule, the marker nucleic acidmolecule comprising a nucleic acid molecule that is linked to a gene,the gene specifically hybridizes to a nucleic acid molecule having anucleic acid sequence selected from the group consisting of SEQ ID NO: 1through SEQ ID NO: 3204 or complements thereof and a complementarynucleic acid molecule obtained from the plant, wherein nucleic acidhybridization between the marker nucleic acid molecule and thecomplementary nucleic acid molecule obtained from the plant permits thedetection of a polymorphism whose presence is predictive of a mutationaffecting the level or pattern of the plant methionine pathway enzyme inthe plant; (B) permitting hybridization between the marker nucleic acidmolecule and the complementary nucleic acid molecule obtained from theplant; and (C) detecting the presence of the polymorphism, wherein thedetection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutationin a plant whose presence is predictive of a mutation affecting thelevel or pattern of a plant methionine pathway enzyme comprising thesteps: (A) incubating, under conditions permitting nucleic acidhybridization, a marker nucleic acid molecule, the marker nucleic acidmolecule comprising a nucleic acid molecule that is linked to a gene,the gene specifically hybridizes to a nucleic acid molecule selectedfrom the group consisting of a nucleic acid molecule that encodes amaize or a soybean methionine adenosyltransferase enzyme or complementthereof, a nucleic acid molecule that encodes a maize or a soybeanS-adenosylmethionine decarboxylase enzyme or complement thereof, anucleic acid molecule that encodes a maize or a soybean aspartate kinaseenzyme or complement thereof, a nucleic acid molecule that encodes amaize or a soybean aspartate-semialdehyde dehydrogenase enzyme orcomplement thereof, a nucleic acid molecule that encodes a maize or asoybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereof,a nucleic acid molecule that encodes a maize or a soybean cystathionineβ-lyase enzyme or complement thereof, a nucleic acid molecule thatencodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof, a nucleic acid molecule that encodes amaize or a soybean adenosylhomocyteinase enzyme or complement thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or complement thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or complementthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or complement thereof and acomplementary nucleic acid molecule obtained from the plant, whereinnucleic acid hybridization between the marker nucleic acid molecule andthe complementary nucleic acid molecule obtained from the plant permitsthe detection of a polymorphism whose presence is predictive of amutation affecting the level or pattern of the plant methionine pathwayenzyme in the plant; (B) permitting hybridization between the markernucleic acid molecule and the complementary nucleic acid moleculeobtained from the plant; and (C) detecting the presence of thepolymorphism, wherein the detection of the polymorphism is predictive ofthe mutation.

A method of producing a plant containing an overexpressed proteincomprising: (A) transforming said plant with a functional nucleic acidmolecule, wherein the functional nucleic acid molecule comprises apromoter region, wherein said promoter region is linked to a structuralregion, wherein said structural region has a nucleic acid sequenceselected from group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204wherein said structural region is linked to a 3′ non-translated sequencethat functions in the plant to cause termination of transcription andaddition of polyadenylated ribonucleotides to a 3′ end of a mRNAmolecule; and wherein said functional nucleic acid molecule results inoverexpression of the protein; and (B) growing said transformed plant.

The present invention also provides a method of producing a plantcontaining an overexpressed plant methionine enzyme comprising: (A)transforming the plant with a functional nucleic acid molecule, whereinthe functional nucleic acid molecule comprises a promoter region,wherein the promoter region is linked to a structural region, whereinthe structural region comprises a nucleic acid molecule having a nucleicacid sequence selected from the group consisting of SEQ ID NO: 1 throughSEQ ID NO: 3204 or fragment thereof; wherein the structural region islinked to a 3′ non-translated sequence that functions in the plant tocause termination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of a mRNA molecule; and wherein thefunctional nucleic acid molecule results in overexpression of the plantmethionine pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method of producing a plantcontaining an overexpressed plant methionine pathway enzyme comprising:(A) transforming the plant with a functional nucleic acid molecule,wherein the functional nucleic acid molecule comprises a promoterregion, wherein the promoter region is linked to a structural region,wherein the structural region comprises a nucleic acid molecule selectedfrom the group consisting of a nucleic acid molecule that encodes amaize or a soybean methionine adenosyltransferase enzyme or fragmentthereof, a nucleic acid molecule that encodes a maize or a soybeanS-adenosylmethionine decarboxylase enzyme or fragment thereof, a nucleicacid molecule that encodes a maize or a soybean aspartate kinase enzymeor fragment thereof, a nucleic acid molecule that encodes a maize or asoybean aspartate-semialdehyde dehydrogenase enzyme or fragment thereof,a nucleic acid molecule that encodes a maize or a soybeanO-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, a nucleicacid molecule that encodes a maize or a soybean cystathionine β-lyaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean adenosylhomocysteinase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; wherein thestructural region is linked to a 3′ non-translated sequence thatfunctions in the plant to cause termination of transcription andaddition of polyadenylated ribonucleotides to a 3′ end of a mRNAmolecule; and wherein the functional nucleic acid molecule results inoverexpression of the plant methionine pathway enzyme protein; and (B)growing the transformed plant.

The present invention also provides a method of producing a plantcontaining reduced levels of a plant methionine pathway enzymecomprising: (A) transforming the plant with a functional nucleic acidmolecule, wherein the functional nucleic acid molecule comprises apromoter region, wherein the promoter region is linked to a structuralregion, wherein the structural region comprises a nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of SEQID NO: 1 through SEQ ID NO: 3204; wherein the structural region islinked to a 3′ non-translated sequence that functions in the plant tocause termination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of a mRNA molecule; and wherein thefunctional nucleic acid molecule results in co-suppression of the plantmethionine pathway enzyme protein; and (B) growing the transformedplant.

The present invention also provides a method of producing a plantcontaining reduced levels of a plant methionine pathway enzymecomprising: (A) transforming the plant with a functional nucleic acidmolecule, wherein the functional nucleic acid molecule comprises apromoter region, wherein the promoter region is linked to a structuralregion, wherein the structural region comprises a nucleic acid moleculehaving a nucleic acid sequence selected from the group consisting of anucleic acid molecule that encodes a maize or a soybean methionineadenosyltransferase enzyme or fragment thereof, a nucleic acid moleculethat encodes a maize or a soybean S-adenosylmethionine decarboxylaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean aspartate kinase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean aspartate-semialdehydedehydrogenase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzymeor fragment thereof, a nucleic acid molecule that encodes a maize or asoybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean adenosylhomocysteinase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; wherein thestructural region is linked to a 3′ non-translated sequence thatfunctions in the plant to cause termination of transcription andaddition of polyadenylated ribonucleotides to a 3′ end of a mRNAmolecule; and wherein the functional nucleic acid molecule results inco-suppression of the plant methionine pathway enzyme; and (B) growingthe transformed plant.

The present invention also provides a method for reducing expression ofa plant methionine pathway enzyme in a plant comprising: (A)transforming the plant with a nucleic acid molecule, the nucleic acidmolecule having an exogenous promoter region which functions in a plantcell to cause the production of a mRNA molecule, wherein the exogenouspromoter region is linked to a transcribed nucleic acid molecule havinga transcribed strand and a non-transcribed strand, wherein thetranscribed strand is complementary to a nucleic acid molecule having anucleic acid sequence selected from the group consisting of SEQ ID NO: 1through SEQ ID NO: 3204 or complements thereof or fragments of eitherand the transcribed strand is complementary to an endogenous mRNAmolecule; and wherein the transcribed nucleic acid molecule is linked toa 3′ non-translated sequence that functions in the plant cell to causetermination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of a mRNA molecule; and (B) growing thetransformed plant.

The present invention also provides a method for reducing expression ofa plant methionine pathway enzyme in a plant comprising: (A)transforming the plant with a nucleic acid molecule, the nucleic acidmolecule having an exogenous promoter region which functions in a plantcell to cause the production of a mRNA molecule, wherein the exogenouspromoter region is linked to a transcribed nucleic acid molecule havinga transcribed strand and a non-transcribed strand, wherein a transcribedmRNA of the transcribed strand is complementary to a nucleic acidmolecule selected from the group consisting of an endogenous mRNAmolecule that encodes a maize or a soybean methionineadenosyltransferase enzyme or fragment thereof, an endogenous mRNAmolecule that encodes a maize or a soybean S-adenosylmethioninedecarboxylase enzyme or fragment thereof, an endogenous mRNA moleculethat encodes a maize or a soybean aspartate kinase enzyme or fragmentthereof, an endogenous mRNA molecule that encodes a maize or a soybeanaspartate-semialdehyde dehydrogenase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybeanO-succinylhomoserine (thiol)-lyase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybean cystathionineβ-lyase enzyme or fragment thereof, an endogenous mRNA molecule thatencodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferaseenzyme or fragment thereof, an endogenous mRNA molecule that encodes amaize or a soybean adenosylhomocysteinase enzyme or fragment thereof, anendogenous mRNA molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, an endogenous mRNA molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and an endogenous mRNA molecule that encodes a maize or asoybean O-acetylhomoserine (thiol)-lyase enzyme or fragment thereof; andwherein the transcribed nucleic acid molecule is linked to a 3′non-translated sequence that functions in the plant cell to causetermination of transcription and addition of polyadenylatedribonucleotides to a 3′ end of a mRNA molecule; and (B) growing thetransformed plant.

The present invention also provides a method of determining anassociation between a polymorphism and a plant trait comprising: (A)hybridizing a nucleic acid molecule specific for the polymorphism togenetic material of a plant, wherein the nucleic acid molecule has anucleic acid sequence selected from the group consisting of SEQ ID NO: 1through SEQ ID NO: 3204 or complements thereof or fragment thereof; and(B) calculating the degree of association between the polymorphism andthe plant trait.

The present invention also provides a method of determining anassociation between a polymorphism and a plant trait comprising: (A)hybridizing a nucleic acid molecule specific for the polymorphism togenetic material of a plant, wherein the nucleic acid molecule isselected from the group consisting of a nucleic acid molecule thatencodes a maize or a soybean methionine adenosyltransferase enzymecomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean S-adenosylmethionine decarboxylase enzymecomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean aspartate kinase enzyme complement thereofor fragment of either, a nucleic acid molecule that encodes a maize or asoybean aspartate-semialdehyde dehydrogenase enzyme complement thereofor fragment of either, a nucleic acid molecule that encodes a maize or asoybean O-succinylhomoserine (thiol)-lyase enzyme complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean cystathionine β-lyase enzyme complement thereof or fragment ofeither, a nucleic acid molecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof or fragment of either; a nucleic acidmolecule that encodes a maize or a soybean adenosylhomocysteinase enzymeor complement thereof or fragment of either, a nucleic acid moleculethat encodes a maize or a soybean cystathionine β-synthase enzyme orcomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean cytsathionine γ-lyase enzyme or complementthereof or fragment of either and a nucleic acid molecule that encodes amaize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complementthereof or fragment of either and (B) calculating the degree ofassociation between the polymorphism and the plant trait.

The present invention also provides a method of isolating a nucleic acidthat encodes a plant methionine pathway enzyme or fragment thereofcomprising: (A) incubating under conditions permitting nucleic acidhybridization, a first nucleic acid molecule comprising a nucleic acidsequence selected from the group consisting of SEQ ID NO: 1 through SEQID NO: 3204 or complements thereof or fragment of either with acomplementary second nucleic acid molecule obtained from a plant cell orplant tissue; (B) permitting hybridization between the first nucleicacid molecule and the second nucleic acid molecule obtained from theplant cell or plant tissue; and (C) isolating the second nucleic acidmolecule.

The present invention also provides a method of isolating a nucleic acidmolecule that encodes a plant methionine pathway enzyme or fragmentthereof comprising: (A) incubating under conditions permitting nucleicacid hybridization, a first nucleic acid molecule selected from thegroup consisting of a nucleic acid molecule that encodes a maize or asoybean methionine adenosyltransferase enzyme complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean S-adenosylmethionine decarboxylase enzyme or complement thereofor fragment of either, a nucleic acid molecule that encodes a maize or asoybean aspartate kinase enzyme complement thereof or fragment ofeither, a nucleic acid molecule that encodes a maize or a soybeanaspartate-semialdehyde dehydrogenase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean O-succinylhomoserine (thiol)-lyase enzyme or complement thereofor fragment of either and a nucleic acid molecule that encodes a maizeor a soybean cystathionine β-lyase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize or asoybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or a soybean adenosylhomocysteinase enzymeor complement thereof or fragment of either, a nucleic acid moleculethat encodes a maize or a soybean cystathionine β-synthase enzyme orcomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean cytsathionine γ-lyase enzyme or complementthereof or fragment of either and a nucleic acid molecule that encodes amaize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complementthereof or fragment of either, with a complementary second nucleic acidmolecule obtained from a plant cell or plant tissue; (B) permittinghybridization between the plant methionine pathway nucleic acid moleculeand the complementary nucleic acid molecule obtained from the plant cellor plant tissue; and (C) isolating the second nucleic acid molecule.

DETAILED DESCRIPTION OF THE INVENTION

Definitions and Agents of the Present Invention

Definitions:

As used herein, a methionine pathway enzyme is any enzyme that isassociated with the synthesis or degradation of methionine.

As used herein, a methionine synthesis enzyme is any enzyme that isassociated with the synthesis of methionine.

As used herein, a methionine degradation enzyme is any enzyme that isassociated with the degradation of methionine.

As used herein, methionine adenosyltransferase is any enzyme thatcatalyzes the conversion of methionine to S-adenosylmethionine.

As used herein, S-adenosylmethionine decarboxylase is any enzyme thatcatalyzes the reaction that converts S-adenosylmethionine to(5-deoxy-5-adenosyl)(3-aminopropyl)methylsulfonuim salt.

As used herein, aspartate kinase is any enzyme that catalyzes theconversion of aspartate to β-aspartyl phosphate.

As used herein, aspartate semialdehyde dehydrogenase is any enzyme thatcatalyzes the conversion of β-aspartyl phosphate toaspartate-semialdehyde via an NADPH-dependent reaction.

As used herein, O-succinylhomoserine (thiol)-lyase refers to any enzymethat catalyzes the conversion of O-phosphohomoserine to and cysteine tocystathionine.

As used herein, cystathionine β-lyase is any enzyme that catalyzes theconversion of cystathionine to homocysteine, pyruvate and ammonia.

As used herein, 5-methyltetrhydropterolytriglutamate-homocysteineS-methyltransferase refers to any enzyme which catalyzes the conversionof homocysteine via methylation to methionine.

As used herein, adenosylhomocysteinase refers to any enzyme thatcatalyzes the ATP-dependent conversion of S-adenosylmethionine (AdoMet)to methylthioadenosine and L-homoserine.

As used herein, cystathionine β-synthase refers to any enzyme thatcatalyzes the conversion of homocysteine and serine to cystathionine.

As used herein, cystathionine γ-lyase refers to any enzyme thatcatalyzes the γ cleavage of cystationine.

As used herein, O-acetyhomoserine (thiol)-lyase refers to any enzymethat catalyzes the conversion of O-acetylhomoserine and sulfur tohomocysteine.

Agents

(a) Nucleic Acid Molecules

Agents of the present invention include plant nucleic acid molecules andmore specifically include maize and soybean nucleic acid molecules andmore specifically include nucleic acid molecules of the maize genotypesB73 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), B73×Mo17(Illinois Foundation Seeds, Champaign, Ill. U.S.A.), DK604 (DekalbGenetics, Dekalb, Ill. U.S.A.), H99 (Illinois Foundation Seeds,Champaign, Ill. U.S.A.), RX601 (Asgrow Seed Company, Des Moines, Iowa),Mo17 (Illinois Foundation Seeds, Champaign, Ill. U.S.A.), and soybeantypes Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa), C1944 (UnitedStates Department of Agriculture (USDA) Soybean Germplasm Collection,Urbana, Ill. U.S.A.), Cristalina (USDA Soybean Germplasm Collection,Urbana, Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA SoybeanGermplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (TohokuUniversity, Morioka, Japan), PI507354 (USDA Soybean GermplasmCollection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed Company, DesMoines, Iowa U.S.A.), PI227687 (USDA Soybean Germplasm Collection,Urbana, Ill. U.S.A.), PI229358 (USDA Soybean Germplasm Collection,Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed Company, Des Moines,Iowa U.S.A.).

A subset of the nucleic acid molecules of the present invention includesnucleic acid molecules that are marker molecules. Another subset of thenucleic acid molecules of the present invention include nucleic acidmolecules that encode a protein or fragment thereof. Another subset ofthe nucleic acid molecules of the present invention are EST molecules.

Fragment nucleic acid molecules may encode significant portion(s) of, orindeed most of, these nucleic acid molecules. Alternatively, thefragments may comprise smaller oligonucleotides (having from about 15 toabout 250 nucleotide residues and more preferably, about 15 to about 30nucleotide residues).

As used herein, an agent, be it a naturally occurring molecule orotherwise may be “substantially purified,” if desired, such that one ormore molecules that is or may be present in a naturally occurringpreparation containing that molecule will have been removed or will bepresent at a lower concentration than that at which it would normally befound.

The agents of the present invention will preferably be “biologicallyactive” with respect to either a structural attribute, such as thecapacity of a nucleic acid to hybridize to another nucleic acidmolecule, or the ability of a protein to be bound by an antibody (or tocompete with another molecule for such binding). Alternatively, such anattribute may be catalytic and thus involve the capacity of the agent tomediate a chemical reaction or response.

The agents of the present invention may also be recombinant. As usedherein, the term recombinant means any agent (e.g. DNA, peptide etc.),that is, or results, however indirect, from human manipulation of anucleic acid molecule.

It is understood that the agents of the present invention may be labeledwith reagents that facilitate detection of the agent (e.g. fluorescentlabels, Prober et al., Science 238:336-340 (1987); Albarella et al., EP144914; chemical labels, Sheldon et al., U.S. Pat. No. 4,582,789;Albarella et al., U.S. Pat. No. 4,563,417; modified bases, Miyoshi etal., EP 119448, all of which are hereby incorporated by reference intheir entirety).

It is further understood, that the present invention providesrecombinant bacterial, mammalian, microbial, insect, fungal and plantcells and viral constructs comprising the agents of the presentinvention. (See, for example, Uses of the Agents of the Invention,Section (a) Plant Constructs and Plant Transformants; Section (b) FungalConstructs and Fungal Transformants; Section (c) Mammalian Constructsand Transformed Mammalian Cells; Section (d) Insect Constructs andTransformed Insect Cells; and Section (e) Bacterial Constructs andTransformed Bacterial Cells)

Nucleic acid molecules or fragments thereof of the present invention arecapable of specifically hybridizing to other nucleic acid moleculesunder certain circumstances. As used herein, two nucleic acid moleculesare said to be capable of specifically hybridizing to one another if thetwo molecules are capable of forming an anti-parallel, double-strandednucleic acid structure. A nucleic acid molecule is said to be the“complement” of another nucleic acid molecule if they exhibit completecomplementarity. As used herein, molecules are said to exhibit “completecomplementarity” when every nucleotide of one of the molecules iscomplementary to a nucleotide of the other. Two molecules are said to be“minimally complementary” if they can hybridize to one another withsufficient stability to permit them to remain annealed to one anotherunder at least conventional “low-stringency” conditions. Similarly, themolecules are said to be “complementary” if they can hybridize to oneanother with sufficient stability to permit them to remain annealed toone another under conventional “high-stringency” conditions.Conventional stringency conditions are described by Sambrook et al.,Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring HarborPress, Cold Spring Harbor, N.Y. (1989) and by Haymes et al., NucleicAcid Hybridization, A Practical Approach, IRL Press, Washington, D.C.(1985), the entirety of which is herein incorporated by reference.Departures from complete complementarity are therefore permissible, aslong as such departures do not completely preclude the capacity of themolecules to form a double-stranded structure. Thus, in order for anucleic acid molecule to serve as a primer or probe it need only besufficiently complementary in sequence to be able to form a stabledouble-stranded structure under the particular solvent and saltconcentrations employed.

Appropriate stringency conditions which promote DNA hybridization, forexample, 6.0× sodium chloride/sodium citrate (SSC) at about 45° C.,followed by a wash of 2.0×SSC at 50° C., are known to those skilled inthe art or can be found in Current Protocols in Molecular Biology, JohnWiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the saltconcentration in the wash step can be selected from a low stringency ofabout 2.0×SSC at 50° C. to a high stringency of about 0.2×SSC at 50° C.In addition, the temperature in the wash step can be increased from lowstringency conditions at room temperature, about 22° C., to highstringency conditions at about 65° C. Both temperature and salt may bevaried, or either the temperature or the salt concentration may be heldconstant while the other variable is changed.

In a preferred embodiment, a nucleic acid of the present invention willspecifically hybridize to one or more of the nucleic acid molecules setforth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereofunder moderately stringent conditions, for example at about 2.0×SSC andabout 65° C.

In a particularly preferred embodiment, a nucleic acid of the presentinvention will include those nucleic acid molecules that specificallyhybridize to one or more of the nucleic acid molecules set forth in SEQID NO: 1 through SEQ ID NO: 3204 or complements thereof under highstringency conditions such as 0.2×SSC and about 65° C.

In one aspect of the present invention, the nucleic acid molecules ofthe present invention have one or more of the nucleic acid sequences setforth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. Inanother aspect of the present invention, one or more of the nucleic acidmolecules of the present invention share between 100% and 90% sequenceidentity with one or more of the nucleic acid sequences set forth in SEQID NO: 1 through SEQ ID NO: 3204 or complements thereof. In a furtheraspect of the present invention, one or more of the nucleic acidmolecules of the present invention share between 100% and 95% sequenceidentity with one or more of the nucleic acid sequences set forth in SEQID NO: 1 through SEQ ID NO: 3204 or complements thereof. In a morepreferred aspect of the present invention, one or more of the nucleicacid molecules of the present invention share between 100% and 98%sequence identity with one or more of the nucleic acid sequences setforth in SEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof. Inan even more preferred aspect of the present invention, one or more ofthe nucleic acid molecules of the present invention share between 100%and 99% sequence identity with one or more of the sequences set forth inSEQ ID NO: 1 through SEQ ID NO: 3204 or complements thereof.

In a further more preferred aspect of the present invention, one or moreof the nucleic acid molecules of the present invention exhibit 100%sequence identity with a nucleic acid molecule present within MONN01,SATMON001 through SATMON031, SATMON033, SATMON034, SATMON˜001,SATMONN01, SATMONN04 through SATMONN006, CMz029 through CMz031, CMz033,CMz035 through CMz037, CMz039 through CMz042, CMz044 through CMz045,CMz047 through CMz050, SOYMON001 through SOYMON038, Soy51 through Soy56,Soy58 through Soy62, Soy65 through Soy66, Soy 68 through Soy73 and Soy76through Soy77, Lib9, Lib22 through Lib25, Lib35, Lib80 through Lib81,Lib 144, Lib146, Lib147, Lib190, Lib3032 through Lib3036 and Lib3099(Monsanto Company, St. Louis, Mo. U.S.A.).

(i) Nucleic Acid Molecules Encoding Proteins or Fragments Thereof

Nucleic acid molecules of the present invention can comprise sequencesthat encode a methionine pathway protein or fragment thereof. Suchproteins or fragments thereof include homologues of known proteins inother organisms.

In a preferred embodiment of the present invention, a maize or soybeanprotein homologue or fragment thereof of the present invention is ahomologue of another plant protein. In another preferred embodiment ofthe present invention, a maize or soybean protein homologue or fragmentthereof of the present invention is a homologue of a fungal protein. Inanother preferred embodiment of the present invention, a maize orsoybean protein homologue of the present invention is a homologue ofmammalian protein. In another preferred embodiment of the presentinvention, a maize or soybean protein homologue or fragment thereof ofthe present invention is a homologue of a bacterial protein. In anotherpreferred embodiment of the present invention, a soybean proteinhomologue or fragment thereof of the present invention is a homologue ofa maize protein. In another preferred embodiment of the presentinvention, a maize protein homologue or fragment thereof of the presentinvention is a homologue of a soybean protein.

In a preferred embodiment of the present invention, the nucleic moleculeof the present invention encodes a maize or soybean homologue protein orfragment thereof where a maize or soybean homologue protein exhibits aBLAST probability score of greater than 1E-12, preferably a BLASTprobability score of between about 1E-30 and about 1E-12, even morepreferably a BLAST probability score of greater than 1E-30 with itshomologue.

In another preferred embodiment of the present invention, the nucleicacid molecule encoding a maize or soybean protein homologue or fragmentthereof or fragment thereof exhibits a % identity with its homologue ofbetween about 25% and about 40%, more preferably of between about 40 andabout 70%, even more preferably of between about 70% and about 90% andeven more preferably between about 90% and 99%. In another preferredembodiment, of the present invention, a maize or soybean proteinhomologue or fragments thereof exhibits a % identity with its homologueof 100%.

In a preferred embodiment of the present invention, the nucleic moleculeof the present invention encodes a maize or soybean homologue protein orfragment thereof where a maize or soybean homologue protein exhibits aBLAST score of greater than 120, preferably a BLAST score of betweenabout 1450 and about 120, even more preferably a BLAST score of greaterthan 1450 with its homologue.

Nucleic acid molecules of the present invention also include non-maize,non-soybean homologues. Preferred non-homologues are selected from thegroup consisting of alfalfa, Arabidopsis, barley, Brassica, broccoli,cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax,an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum,strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir,eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses,sunflower, oil palm and Phaseolus.

In a preferred embodiment, nucleic acid molecules having SEQ ID NO: 1through SEQ ID NO: 3204 or complements and fragments of either can beutilized to obtain such homologues.

The degeneracy of the genetic code, which allows different nucleic acidsequences to code for the same protein or peptide, is known in theliterature. (U.S. Pat. No. 4,757,006, the entirety of which is hereinincorporated by reference).

In an aspect of the present invention, one or more of the nucleic acidmolecules of the present invention differ in nucleic acid sequence fromthose encoding maize or soybean homologue or fragment thereof in SEQ IDNO: 1 through SEQ ID NO: 3204 due to the degeneracy in the genetic codein that they encode the same protein but differ in nucleic acidsequence.

In another further aspect of the present invention, one or more of thenucleic acid molecules of the present invention differ in nucleic acidsequence from those encoding maize or soybean homologue or fragmentthereof in SEQ ID NO: 1 through SEQ ID NO: 3204 due to fact that thedifferent nucleic acid sequence encodes a protein having one or moreconservative amino acid residue. Examples of conservative substitutionsare set forth in Table 1. It is understood that codons capable of codingfor such conservative substitutions are known in the art. TABLE 1Original Residue Conservative Substitutions Ala Ser Arg Lys Asn Gln; HisAsp Glu Cys Ser; Ala Gln Asn Glu Asp Gly Pro His Asn; Gln Ile Leu; ValLeu Ile; Val Lys Arg; Gln; Glu Met Leu; Ile Phe Met; Leu; Tyr Ser ThrThr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu

In a further aspect of the present invention, one or more of the nucleicacid molecules of the present invention differ in nucleic acid sequencefrom those encoding a maize or soybean homologue or fragment thereof setforth in SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereof due tothe fact that one or more codons encoding an amino acid has beensubstituted for a codon that encodes a nonessential substitution of theamino acid originally encoded.

Agents of the present invention include nucleic acid molecules thatencode a maize or soybean methionine pathway protein or fragment thereofand particularly substantially purified nucleic acid molecules selectedfrom the group consisting of a nucleic acid molecule that encodes amaize or soybean methionine adenosyltransferase protein or fragmentthereof, a nucleic acid molecule that encodes a maize or soybeanS-adenosylmethionine decarboxylase protein or fragment thereof, anucleic acid molecule that encodes a maize or soybean aspartate kinaseprotein or fragment thereof, a nucleic acid molecule that encode a maizeor soybean aspartate-semialdehyde dehydrogenase protein or fragmentthereof, a nucleic acid molecule that encodes a maize or soybeanO-succinylhomoserine (thiol)-lyase protein or fragment thereof, anucleic acid molecule that encodes a maize or soybean cystathionineβ-lyase protein or fragment thereof, a nucleic acid molecule thatencodes a maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseprotein or fragment thereof, a nucleic acid molecule that encodes amaize or soybean adenosylhomocysteine protein or fragment thereof, anucleic acid molecule that encodes a maize or soybean cystathionineβ-synthase protein or fragment thereof, a nucleic acid molecule thatencodes a maize or soybean cystathionine γ-lyase protein or fragmentthereof, and a nucleic acid molecule that encodes a maize or soybeanO-acetylhomoserine (thiol)-lyase protein or fragment thereof.

Non-limiting examples of such nucleic acid molecules of the presentinvention are nucleic acid molecules comprising: SEQ ID NO: 1 throughSEQ ID NO: 3204 or fragment thereof that encode for a methionine pathwayprotein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 429 and SEQID NO: 1635 through SEQ ID NO: 2479 or fragment thereof that encode fora methionine adenosyltransferase protein or fragment thereof, SEQ ID NO:430 through SEQ ID NO: 857 and SEQ ID NO: 2480 through SEQ ID NO: 2623or fragment thereof that encode for a S-adenosylmethionine decarboxylaseprotein or fragment thereof, SEQ ID NO: 858 through SEQ ID NO: 900 andSEQ ID NO: 2624 through SEQ ID NO: 2648 or fragment thereof that encodefor a aspartate kinase protein or fragment thereof, SEQ ID NO: 901through SEQ ID NO: 904 and SEQ ID NO: 2649 through SEQ ID NO: 2654 orfragment thereof that encode for a aspartate-semialdehyde dehydrogenaseprotein or fragment thereof, SEQ ID NO: 905 through SEQ ID NO: 953 andSEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereof that encodefor a O-succinylhomoserine (thiol)-lyase protein or fragment thereof,SEQ ID NO: 954 through SEQ ID NO: 963 and SEQ ID NO: 2655 through SEQ IDNO: 2660 or fragment thereof that encode for a cystathionine β-lyaseprotein or fragment thereof, SEQ ID NO: 964 through SEQ ID NO: 1353 andSEQ ID NO: 2666 through SEQ ID NO: 2992 or fragment thereof that encodefor a5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferaseprotein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO: 1630 andSEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment thereof that encodefor an adenosylhomocysteinase protein or fragment thereof, SEQ ID NO:1631 through SEQ ID NO: 1632 or fragment thereof that encode for acystathionine β-synthase protein or fragment thereof, SEQ ID NO: 1633through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 orfragment thereof that encode for a cystathionine γ-lyase protein orfragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 orfragment thereof that encode for an O-acetylhomoserine (thiol)-lyaseprotein or fragment thereof.

A nucleic acid molecule of the present invention can also encode anhomologue of a maize or soybean methionine adenosyltransferase orfragment thereof, a maize or soybean S-adenosylmethionine decarboxylaseor fragment thereof, a maize or soybean aspartate kinase or fragmentthereof, a maize or soybean aspartate-semialdehyde dehydrogenase orfragment thereof, a maize or soybean O-succinylhomoserine (thiol)-lyaseor fragment thereof, a maize or soybean cystathionine β-lyase orfragment thereof, a maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseor fragment thereof, a maize or soybean adenosylhomocysteinease orfragment thereof, a maize or soybean cystathionine β-synthase orfragment thereof, a maize or soybean cystathionine γ-lyase or fragmentthereof or a maize or soybean O-acetylhomoserine (thiol)-lyase orfragment thereof. As used herein a homologue protein molecule orfragment thereof is a counterpart protein molecule or fragment thereofin a second species (e.g., maize methionine adenosyltransferase proteinis a homologue of Arabidopsis' methionine adenosyltransferase protein).

(ii) Nucleic Acid Molecule Markers and Probes

One aspect of the present invention concerns markers that includenucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 3204 orcomplements thereof or fragments of either that can act as markers.Genetic markers of the present invention include “dominant” or“codominant” markers. “Codominant markers” reveal the presence of two ormore alleles (two per diploid individual) at a locus. “Dominant markers”reveal the presence of only a single allele per locus. The presence ofthe dominant marker phenotype (e.g., a band of DNA) is an indicationthat one allele is present in either the homozygous or heterozygouscondition. The absence of the dominant marker phenotype (e.g. absence ofa DNA band) is merely evidence that “some other” undefined allele ispresent. In the case of populations where individuals are predominantlyhomozygous and loci are predominately dimorphic, dominant and codominantmarkers can be equally valuable. As populations become more heterozygousand multi-allelic, codominant markers often become more informative ofthe genotype than dominant markers. Marker molecules can be, forexample, capable of detecting polymorphisms such as single nucleotidepolymorphisms (SNPs).

SNPs are single base changes in genomic DNA sequence. They occur atgreater frequency and are spaced with a greater uniformly throughout agenome than other reported forms of polymorphism. The greater frequencyand uniformity of SNPs means that there is greater probability that sucha polymorphism will be found near or in a genetic locus of interest thanwould be the case for other polymorphisms. SNPs are located inprotein-coding regions and noncoding regions of a genome. Some of theseSNPs may result in defective or variant protein expression (e.g., as aresults of mutations or defective splicing). Analysis (genotyping) ofcharacterized SNPs can require only a plus/minus assay rather than alengthy measurement, permitting easier automation.

SNPs can be characterized using any of a variety of methods. Suchmethods include the direct or indirect sequencing of the site, the useof restriction enzymes (Botstein et al., Am. J. Hum. Genet. 32:314-331(1980), the entirety of which is herein incorporated reference;Konieczny and Ausubel, Plant J. 4:403-410 (1993), the entirety of whichis herein incorporated by reference), enzymatic and chemical mismatchassays (Myers et al., Nature 313:495-498 (1985), the entirety of whichis herein incorporated by reference), allele-specific PCR (Newton etal., Nucl. Acids Res. 17:2503-2516 (1989), the entirety of which isherein incorporated by reference; Wu et al., Proc. Natl. Acad. Sci.(U.S.A.) 86:2757-2760 (1989), the entirety of which is hereinincorporated by reference), ligase chain reaction (Barany, Proc. Natl.Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is hereinincorporated by reference), single-strand conformation polymorphismanalysis (Labrune et al., Am. J. Hum. Genet. 48: 1115-1120 (1991), theentirety of which is herein incorporated by reference), primer-directednucleotide incorporation assays (Kuppuswami et al., Proc. Natl. Acad.Sci. USA 88:1143-1147 (1991), the entirety of which is hereinincorporated by reference), dideoxy fingerprinting (Sarkar et al.,Genomics 13:441-443 (1992), the entirety of which is herein incorporatedby reference), solid-phase ELISA-based oligonucleotide ligation assays(Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the entirety ofwhich is herein incorporated by reference), oligonucleotidefluorescence-quenching assays (Livak et al., PCR Methods Appl. 4:357-362(1995), the entirety of which is herein incorporated by reference),5′-nuclease allele-specific hybridization TaqMan assay (Livak et al.,Nature Genet. 9:341-342 (1995), the entirety of which is hereinincorporated by reference), template-directed dye-terminatorincorporation (TDI) assay (Chen and Kwok, Nucl. Acids Res. 25:347-353(1997), the entirety of which is herein incorporated by reference),allele-specific molecular beacon assay (Tyagi et al., Nature Biotech.16: 49-53 (1998), the entirety of which is herein incorporated byreference), PinPoint assay (Haff and Smimov, Genome Res. 7: 378-388(1997), the entirety of which is herein incorporated by reference) anddCAPS analysis (Neff et al., Plant J. 14:387-392 (1998), the entirety ofwhich is herein incorporated by reference).

Additional markers, such as AFLP markers, RFLP markers and RAPD markers,can be utilized (Walton, Seed World 22-29 (July, 1993), the entirety ofwhich is herein incorporated by reference; Burow and Blake, MolecularDissection of Complex Traits, 13-29, Paterson (ed.), CRC Press, New York(1988), the entirety of which is herein incorporated by reference). DNAmarkers can be developed from nucleic acid molecules using restrictionendonucleases, the PCR and/or DNA sequence information. RFLP markersresult from single base changes or insertions/deletions. Thesecodominant markers are highly abundant in plant genomes, have a mediumlevel of polymorphism and are developed by a combination of restrictionendonuclease digestion and Southern blotting hybridization. CAPS aresimilarly developed from restriction nuclease digestion but only ofspecific PCR products. These markers are also codominant, have a mediumlevel of polymorphism and are highly abundant in the genome. The CAPSresult from single base changes and insertions/deletions.

Another marker type, RAPDs, are developed from DNA amplification withrandom primers and result from single base changes andinsertions/deletions in plant genomes. They are dominant markers with amedium level of polymorphisms and are highly abundant. AFLP markersrequire using the PCR on a subset of restriction fragments from extendedadapter primers. These markers are both dominant and codominant arehighly abundant in genomes and exhibit a medium level of polymorphism.

SSRs require DNA sequence information. These codominant markers resultfrom repeat length changes, are highly polymorphic and do not exhibit ashigh a degree of abundance in the genome as CAPS, AFLPs and RAPDs SNPsalso require DNA sequence information. These codominant markers resultfrom single base substitutions. They are highly abundant and exhibit amedium of polymorphism (Rafalski et al., In: Nonmammalian GenomicAnalysis, Birren and Lai (ed.), Academic Press, San Diego, Calif., pp.75-134 (1996), the entirety of which is herein incorporated byreference). It is understood that a nucleic acid molecule of the presentinvention may be used as a marker.

A PCR probe is a nucleic acid molecule capable of initiating apolymerase activity while in a double-stranded structure to with anothernucleic acid. Various methods for determining the structure of PCRprobes and PCR techniques exist in the art. Computer generated searchesusing programs such as Primer3(www-genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline(www-genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole etal., BioTechniques 25:112-123 (1998) the entirety of which is hereinincorporated by reference), for example, can be used to identifypotential PCR primers.

It is understood that a fragment of one or more of the nucleic acidmolecules of the present invention may be a probe and specifically a PCRprobe.

(b) Protein and Peptide Molecules

A class of agents comprises one or more of the protein or fragmentsthereof or peptide molecules encoded by SEQ ID NO: 1 through SEQ ID NO:3204 or one or more of the protein or fragment thereof and peptidemolecules encoded by other nucleic acid agents of the present invention.As used herein, the term “protein molecule” or “peptide molecule”includes any molecule that comprises five or more amino acids. It iswell known in the art that proteins may undergo modification, includingpost-translational modifications, such as, but not limited to, disulfidebond formation, glycosylation, phosphorylation, or oligomerization.Thus, as used herein, the term “protein molecule” or “peptide molecule”includes any protein molecule that is modified by any biological ornon-biological process. The terms “amino acid” and “amino acids” referto all naturally occurring L-amino acids. This definition is meant toinclude norleucine, ornithine, homocysteine and homoserine.

Non-limiting examples of the protein or fragment thereof of the presentinvention include a maize or soybean methionine pathway protein orfragment thereof; a maize or soybean methionine adenosyltransferase orfragment thereof, a maize or soybean S-adenosylmethionine decarboxylaseor fragment thereof, a maize or soybean aspartate kinase or fragmentthereof, a maize or soybean aspartate-semialdehyde dehydrogenase orfragment thereof, a maize or soybean O-succinylhomoserine (thiol)-lyaseor fragment thereof, a maize or soybean cystathionine β-lyase orfragment thereof, a maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseor fragment thereof, a maize or soybean adenosylhomocysteinase orfragment thereof, a maize or soybean cystathionine β-synthase orfragment thereof, a maize or soybean cystathionine γ-lyase or fragmentthereof or a maize or soybean O-acetylhomoserine (thiol)-lyase orfragment thereof.

Non-limiting examples of the protein or fragment molecules of thepresent invention are an methionine pathway protein or fragment thereofencoded by: SEQ ID NO: 1 through SEQ ID NO: 3204 or fragment thereofthat encode for a methionine pathway protein or fragment thereof, SEQ IDNO: 1 through SEQ ID NO: 429 and SEQ ID NO: 1635 through SEQ ID NO: 2479or fragment thereof that encode for a methionine adenosyltransferaseprotein or fragment thereof, SEQ ID NO: 430 through SEQ ID NO: 857 andSEQ ID NO: 2480 through SEQ ID NO: 2623 or fragment thereof that encodefor a S-adenosylmethionine decarboxylase protein or fragment thereof,SEQ ID NO: 858 through SEQ ID NO: 900 and SEQ ID NO: 2624 through SEQ IDNO: 2648 or fragment thereof that encode for a aspartate kinase proteinor fragment thereof, SEQ ID NO: 901 through SEQ ID NO: 904 and SEQ IDNO: 2649 through SEQ ID NO: 2654 or fragment thereof that encode for aaspartate-semialdehyde dehydrogenase protein or fragment thereof, SEQ IDNO: 905 through SEQ ID NO: 953 and SEQ ID NO: 2655 through SEQ ID NO:2660 or fragment thereof that encode for a O-succinylhomoserine(thiol)-lyase protein or fragment thereof, SEQ ID NO: 954 through SEQ IDNO: 963 and SEQ ID NO: 2655 through SEQ ID NO: 2660 or fragment thereofthat encode for a cystathionine β-lyase or fragment thereof, SEQ ID NO:964 through SEQ ID NO: 1353 and SEQ ID NO: 2666 through SEQ ID NO: 2992or fragment thereof that encode for a5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseprotein or fragment thereof, SEQ ID NO: 1354 through SEQ ID NO: 1630 andSEQ ID NO: 2993 through SEQ ID NO: 3199 or fragment thereof that encodefor an adenosylhomocysteinase protein or fragment thereof, SEQ ID NO:1631 through SEQ ID NO: 1632 or fragment thereof that encode for acystathionine β-synthase protein or fragment thereof, SEQ ID NO: 1633through SEQ ID NO: 1634 and SEQ ID NO: 3203 through SEQ ID NO: 3204 orfragment thereof that encode for a cystathionine γ-lyase protein orfragment thereof, and SEQ ID NO: 3200 through SEQ ID NO: 3202 orfragment thereof that encode for an O-acetylhomoserine (thiol)-lyaseprotein or fragment thereof.

One or more of the protein or fragment of peptide molecules may beproduced via chemical synthesis, or more preferably, by expressing in asuitable bacterial or eucaryotic host. Suitable methods for expressionare described by Sambrook et al., (In: Molecular Cloning, A LaboratoryManual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.(1989)), or similar texts. For example, the protein may be expressed in,for example, Uses of the Agents of the Invention, Section (a) PlantConstructs and Plant Transformants; Section (b) Fungal Constructs andFungal Transformants; Section (c) Mammalian Constructs and TransformedMammalian Cells; Section (d) Insect Constructs and Transformed InsectCells; and Section (e) Bacterial Constructs and Transformed BacterialCells.

A “protein fragment” is a peptide or polypeptide molecule whose aminoacid sequence comprises a subset of the amino acid sequence of thatprotein. A protein or fragment thereof that comprises one or moreadditional peptide regions not derived from that protein is a “fusion”protein. Such molecules may be derivatized to contain carbohydrate orother moieties (such as keyhole limpet hemocyanin, etc.). Fusion proteinor peptide molecules of the present invention are preferably producedvia recombinant means.

Another class of agents comprise protein or peptide molecules orfragments or fusions thereof encoded by SEQ ID NO: 1 through SEQ ID NO:3204 or complements thereof in which conservative, non-essential ornon-relevant amino acid residues have been added, replaced or deleted.Computerized means for designing modifications in protein structure areknown in the art (Dahiyat and Mayo, Science 278:82-87 (1997), theentirety of which is herein incorporated by reference).

The protein molecules of the present invention include plant homologueproteins. An example of such a homologue is a homologue protein of anon-maize or non soybean plant species, that include but not limited toalfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus,cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamentalplant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry,sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus,apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower,oil palm, Phaseolus etc. Particularly preferred non-maize or non-soybeanfor use for the isolation of homologs would include, Arabidopsis,barley, cotton, oat, oilseed rape, rice, canola, ornamentals, sugarcane,sugarbeet, tomato, potato, wheat and turf grasses. Such a homologue canbe obtained by any of a variety of methods. Most preferably, asindicated above, one or more of the disclosed sequences (SEQ ID NO: 1through SEQ ID NO: 3204 or complements thereof) will be used to define apair of primers that may be used to isolate the homologue-encodingnucleic acid molecules from any desired species. Such molecules can beexpressed to yield homologues by recombinant means.

(c) Antibodies

One aspect of the present invention concerns antibodies, single-chainantigen binding molecules, or other proteins that specifically bind toone or more of the protein or peptide molecules of the present inventionand their homologues, fusions or fragments. Such antibodies may be usedto quantitatively or qualitatively detect the protein or peptidemolecules of the present invention. As used herein, an antibody orpeptide is said to “specifically bind” to a protein or peptide moleculeof the present invention if such binding is not competitively inhibitedby the presence of non-related molecules.

Nucleic acid molecules that encode all or part of the protein of thepresent invention can be expressed, via recombinant means, to yieldprotein or peptides that can in turn be used to elicit antibodies thatare capable of binding the expressed protein or peptide. Such antibodiesmay be used in immunoassays for that protein. Such protein-encodingmolecules, or their fragments may be a “fusion” molecule (i.e., a partof a larger nucleic acid molecule) such that, upon expression, a fusionprotein is produced. It is understood that any of the nucleic acidmolecules of the present invention may be expressed, via recombinantmeans, to yield proteins or peptides encoded by these nucleic acidmolecules.

The antibodies that specifically bind proteins and protein fragments ofthe present invention may be polyclonal or monoclonal and may compriseintact immunoglobulins, or antigen binding portions of immunoglobulinsfragments (such as (F(ab′), F(ab′)₂), or single-chain immunoglobulinsproducible, for example, via recombinant means. It is understood thatpractitioners are familiar with the standard resource materials whichdescribe specific conditions and procedures for the construction,manipulation and isolation of antibodies (see, for example, Harlow andLane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor Press,Cold Spring Harbor, N.Y. (1988), the entirety of which is hereinincorporated by reference).

Murine monoclonal antibodies are particularly preferred. BALB/c mice arepreferred for this purpose, however, equivalent strains may also beused. The animals are preferably immunized with approximately 25 μg ofpurified protein (or fragment thereof) that has been emulsified in asuitable adjuvant (such as TiterMax adjuvant (Vaxcel, Norcross, Ga.)).Immunization is preferably conducted at two intramuscular sites, oneintraperitoneal site and one subcutaneous site at the base of the tail.An additional i.v. injection of approximately 25 μg of antigen ispreferably given in normal saline three weeks later. After approximately11 days following the second injection, the mice may be bled and theblood screened for the presence of anti-protein or peptide antibodies.Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) isemployed for this purpose.

More preferably, the mouse having the highest antibody titer is given athird i.v. injection of approximately 25 μg of the same protein orfragment. The splenic leukocytes from this animal may be recovered 3days later and then permitted to fuse, most preferably, usingpolyethylene glycol, with cells of a suitable myeloma cell line (suchas, for example, the P3X63Ag8.653 myeloma cell line). Hybridoma cellsare selected by culturing the cells under “HAT”(hypoxanthine-aminopterin-thymine) selection for about one week. Theresulting clones may then be screened for their capacity to producemonoclonal antibodies (“mAbs”), preferably by direct ELISA.

In one embodiment, anti-protein or peptide monoclonal antibodies areisolated using a fusion of a protein or peptide of the presentinvention, or conjugate of a protein or peptide of the presentinvention, as immunogens. Thus, for example, a group of mice can beimmunized using a fusion protein emulsified in Freund's completeadjuvant (e.g. approximately 50 μg of antigen per immunization). Atthree week intervals, an identical amount of antigen is emulsified inFreund's incomplete adjuvant and used to immunize the animals. Ten daysfollowing the third immunization, serum samples are taken and evaluatedfor the presence of antibody. If antibody titers are too low, a fourthbooster can be employed. Polysera capable of binding the protein orpeptide can also be obtained using this method.

In a preferred procedure for obtaining monoclonal antibodies, thespleens of the above-described immunized mice are removed, disrupted andimmune splenocytes are isolated over a ficoll gradient. The isolatedsplenocytes are fused, using polyethylene glycol with BALB/c-derivedHGPRT (hypoxanthine guanine phosphoribosyl transferase) deficientP3x63xAg8.653 plasmacytoma cells. The fused cells are plated into 96well microtiter plates and screened for hybridoma fusion cells by theircapacity to grow in culture medium supplemented with hypothanthine,aminopterin and thymidine for approximately 2-3 weeks.

Hybridoma cells that arise from such incubation are preferably screenedfor their capacity to produce an immunoglobulin that binds to a proteinof interest. An indirect ELISA may be used for this purpose. In brief,the supernatants of hybridomas are incubated in microtiter wells thatcontain immobilized protein. After washing, the titer of boundimmunoglobulin can be determined using, for example, a goat anti-mouseantibody conjugated to horseradish peroxidase. After additional washing,the amount of immobilized enzyme is determined (for example through theuse of a chromogenic substrate). Such screening is performed as quicklyas possible after the identification of the hybridoma in order to ensurethat a desired clone is not overgrown by non-secreting neighbor cells.Desirably, the fusion plates are screened several times since the ratesof hybridoma growth vary. In a preferred sub-embodiment, a differentantigenic form may be used to screen the hybridoma. Thus, for example,the splenocytes may be immunized with one immunogen, but the resultinghybridomas can be screened using a different immunogen. It is understoodthat any of the protein or peptide molecules of the present inventionmay be used to raise antibodies.

As discussed below, such antibody molecules or their fragments may beused for diagnostic purposes. Where the antibodies are intended fordiagnostic purposes, it may be desirable to derivatize them, for examplewith a ligand group (such as biotin) or a detectable marker group (suchas a fluorescent group, a radioisotope or an enzyme).

The ability to produce antibodies that bind the protein or peptidemolecules of the present invention permits the identification of mimeticcompounds of those molecules. A “mimetic compound” is a compound that isnot that compound, or a fragment of that compound, but which nonethelessexhibits an ability to specifically bind to antibodies directed againstthat compound.

It is understood that any of the agents of the present invention can besubstantially purified and/or be biologically active and/or recombinant.

Uses of the Agents of the Invention

Nucleic acid molecules and fragments thereof of the present inventionmay be employed to obtain other nucleic acid molecules from the samespecies (e.g., ESTs or fragment thereof from maize may be utilized toobtain other nucleic acid molecules from maize). Such nucleic acidmolecules include the nucleic acid molecules that encode the completecoding sequence of a protein and promoters and flanking sequences ofsuch molecules. In addition, such nucleic acid molecules include nucleicacid molecules that encode for other isozymes or gene family members.Such molecules can be readily obtained by using the above-describednucleic acid molecules or fragments thereof to screen cDNA or genomiclibraries obtained from maize or soybean. Methods for forming suchlibraries are well known in the art.

Nucleic acid molecules and fragments thereof of the present inventionmay also be employed to obtain nucleic acid homologues. Such homologuesinclude the nucleic acid molecule of other plants or other organisms(e.g., alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage,citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, anornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum,strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir,eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses,sunflower, oil palm, Phaseolus, etc.) including the nucleic acidmolecules that encode, in whole or in part, protein homologues of otherplant species or other organisms, sequences of genetic elements such aspromoters and transcriptional regulatory elements. Such molecules can bereadily obtained by using the above-described nucleic acid molecules orfragments thereof to screen cDNA or genomic libraries obtained from suchplant species. Methods for forming such libraries are well known in theart. Such homologue molecules may differ in their nucleotide sequencesfrom those found in one or more of SEQ ID NO: 1 through SEQ ID NO: 3204or complements thereof because complete complementarity is not neededfor stable hybridization. The nucleic acid molecules of the presentinvention therefore also include molecules that, although capable ofspecifically hybridizing with the nucleic acid molecules may lack“complete complementarity.”

Any of a variety of methods may be used to obtain one or more of theabove-described nucleic acid molecules (Zamechik et al., Proc. Natl.Acad. Sci. (U.S.A.) 83:4143-4146 (1986), the entirety of which is hereinincorporated by reference; Goodchild et al., Proc. Natl. Acad. Sci.(U.S.A.) 85:5507-5511 (1988), the entirety of which is hereinincorporated by reference; Wickstrom et al., Proc. Natl. Acad. Sci.(U.S.A.) 85:1028-1032 (1988), the entirety of which is hereinincorporated by reference; Holt et al., Molec. Cell. Biol. 8:963-973(1988), the entirety of which is herein incorporated by reference;Gerwirtz et al., Science 242:1303-1306 (1988), the entirety of which isherein incorporated by reference; Anfossi et al., Proc. Natl. Acad. Sci.(U.S.A.) 86:3379-3383 (1989), the entirety of which is hereinincorporated by reference; Becker et al., EMBO J. 8:3685-3691 (1989);the entirety of which is herein incorporated by reference). Automatednucleic acid synthesizers may be employed for this purpose. In lieu ofsuch synthesis, the disclosed nucleic acid molecules may be used todefine a pair of primers that can be used with the polymerase chainreaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol.51:263-273 (1986); Erlich et al., European Patent 50,424; EuropeanPatent 84,796; European Patent 258,017; European Patent 237,362; Mullis,European Patent 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich,U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194, allof which are herein incorporated by reference in their entirety) toamplify and obtain any desired nucleic acid molecule or fragment.

Promoter sequence(s) and other genetic elements, including but notlimited to transcriptional regulatory flanking sequences, associatedwith one or more of the disclosed nucleic acid sequences can also beobtained using the disclosed nucleic acid sequence provided herein. Inone embodiment, such sequences are obtained by incubating EST nucleicacid molecules or preferably fragments thereof with members of genomiclibraries (e.g. maize and soybean) and recovering clones that hybridizeto the EST nucleic acid molecule or fragment thereof. In a secondembodiment, methods of “chromosome walking,” or inverse PCR may be usedto obtain such sequences (Frohman et al., Proc. Natl. Acad. Sci.(U.S.A.) 85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci.(U.S.A.) 86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048(1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et al.,Method Mol. Biol. 67:287-294 (1997); Benkel et al., Genet. Anal.13:123-127 (1996); Hartl et al., Methods Mol. Biol. 58:293-301 (1996),all of which are herein incorporated by reference in their entirety).

The nucleic acid molecules of the present invention may be used toisolate promoters of cell enhanced, cell specific, tissue enhanced,tissue specific, developmentally or environmentally regulated expressionprofiles. Isolation and functional analysis of the 5′ flanking promotersequences of these genes from genomic libraries, for example, usinggenomic screening methods and PCR techniques would result in theisolation of useful promoters and transcriptional regulatory elements.These methods are known to those of skill in the art and have beendescribed (See, for example, Birren et al., Genome Analysis: AnalyzingDNA, 1, (1997), Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y., the entirety of which is herein incorporated by reference).Promoters obtained utilizing the nucleic acid molecules of the presentinvention could also be modified to affect their controlcharacteristics. Examples of such modifications would include but arenot limited to enhanced sequences as reported in Uses of the Agents ofthe Invention, Section (a) Plant Constructs and Plant Transformants.Such genetic elements could be used to enhance gene expression of newand existing traits for crop improvements.

In one sub-aspect, such an analysis is conducted by determining thepresence and/or identity of polymorphism(s) by one or more of thenucleic acid molecules of the present invention and more preferably oneor more of the EST nucleic acid molecule or fragment thereof which areassociated with a phenotype, or a predisposition to that phenotype.

Any of a variety of molecules can be used to identify suchpolymorphism(s). In one embodiment, one or more of the EST nucleic acidmolecules (or a sub-fragment thereof) may be employed as a markernucleic acid molecule to identify such polymorphism(s). Alternatively,such polymorphisms can be detected through the use of a marker nucleicacid molecule or a marker protein that is genetically linked to (i.e., apolynucleotide that co-segregates with) such polymorphism(s).

In an alternative embodiment, such polymorphisms can be detected throughthe use of a marker nucleic acid molecule that is physically linked tosuch polymorphism(s). For this purpose, marker nucleic acid moleculescomprising a nucleotide sequence of a polynucleotide located within 1 mbof the polymorphism(s) and more preferably within 100 kb of thepolymorphism(s) and most preferably within 10 kb of the polymorphism(s)can be employed.

The genomes of animals and plants naturally undergo spontaneous mutationin the course of their continuing evolution (Gusella, Ann. Rev. Biochem.55:831-854 (1986)). A “polymorphism” is a variation or difference in thesequence of the gene or its flanking regions that arises in some of themembers of a species. The variant sequence and the “original” sequenceco-exist in the species' population. In some instances, suchco-existence is in stable or quasi-stable equilibrium.

A polymorphism is thus said to be “allelic,” in that, due to theexistence of the polymorphism, some members of a species may have theoriginal sequence (i.e., the original “allele”) whereas other membersmay have the variant sequence (i.e., the variant “allele”). In thesimplest case, only one variant sequence may exist and the polymorphismis thus said to be di-allelic. In other cases, the species' populationmay contain multiple alleles and the polymorphism is termed tri-allelic,etc. A single gene may have multiple different unrelated polymorphisms.For example, it may have a di-allelic polymorphism at one site and amulti-allelic polymorphism at another site.

The variation that defines the polymorphism may range from a singlenucleotide variation to the insertion or deletion of extended regionswithin a gene. In some cases, the DNA sequence variations are in regionsof the genome that are characterized by short tandem repeats (STRs) thatinclude tandem di- or tri-nucleotide repeated motifs of nucleotides.Polymorphisms characterized by such tandem repeats are referred to as“variable number tandem repeat” (“VNTR”) polymorphisms. VNTRs have beenused in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour etal., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol.39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003;Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat. No.5,175,082; Jeffreys et al., Amer. J. Hum. Genet. 39:11-24 (1986);Jeffreys et al., Nature 316:76-79 (1985); Gray et al., Proc. R. Acad.Soc. Lond. 243:241-253 (1991); Moore et al., Genomics 10:654-660 (1991);Jeffreys et al., Anim. Genet. 18:1-15 (1987); Hillel et al., Anim.Genet. 20:145-155 (1989); Hillel et al., Genet. 124:783-789 (1990), allof which are herein incorporated by reference in their entirety).

The detection of polymorphic sites in a sample of DNA may be facilitatedthrough the use of nucleic acid amplification methods. Such methodsspecifically increase the concentration of polynucleotides that span thepolymorphic site, or include that site and sequences located eitherdistal or proximal to it. Such amplified molecules can be readilydetected by gel electrophoresis or other means.

The most preferred method of achieving such amplification employs thepolymerase chain reaction (“PCR”) (Mullis et al., Cold Spring HarborSymp. Quant. Biol. 51:263-273 (1986); Erlich et al., European PatentAppln. 50,424; European Patent Appln. 84,796; European PatentApplication 258,017; European Patent Appln. 237,362; Mullis, EuropeanPatent Appln. 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich,U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194),using primer pairs that are capable of hybridizing to the proximalsequences that define a polymorphism in its double-stranded form.

In lieu of PCR, alternative methods, such as the “Ligase Chain Reaction”(“LCR”) may be used (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193(1991), the entirety of which is herein incorporated by reference). LCRuses two pairs of oligonucleotide probes to exponentially amplify aspecific target. The sequences of each pair of oligonucleotides isselected to permit the pair to hybridize to abutting sequences of thesame strand of the target. Such hybridization forms a substrate for atemplate-dependent ligase. As with PCR, the resulting products thusserve as a template in subsequent cycles and an exponentialamplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal anddistal sequences of the same strand of a polymorphic site. In oneembodiment, either oligonucleotide will be designed to include theactual polymorphic site of the polymorphism. In such an embodiment, thereaction conditions are selected such that the oligonucleotides can beligated together only if the target molecule either contains or lacksthe specific nucleotide that is complementary to the polymorphic sitepresent on the oligonucleotide. Alternatively, the oligonucleotides maybe selected such that they do not include the polymorphic site (see,Segev, PCT Application WO 90/01069, the entirety of which is hereinincorporated by reference).

The “Oligonucleotide Ligation Assay” (“OLA”) may alternatively beemployed (Landegren et al., Science 241:1077-1080 (1988), the entiretyof which is herein incorporated by reference). The OLA protocol uses twooligonucleotides which are designed to be capable of hybridizing toabutting sequences of a single strand of a target. OLA, like LCR, isparticularly suited for the detection of point mutations. Unlike LCR,however, OLA results in “linear” rather than exponential amplificationof the target sequence.

Nickerson et al., have described a nucleic acid detection assay thatcombines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad.Sci. (U.S.A.) 87:8923-8927 (1990), the entirety of which is hereinincorporated by reference). In this method, PCR is used to achieve theexponential amplification of target DNA, which is then detected usingOLA. In addition to requiring multiple and separate, processing steps,one problem associated with such combinations is that they inherit allof the problems associated with PCR and OLA.

Schemes based on ligation of two (or more) oligonucleotides in thepresence of nucleic acid having the sequence of the resulting“di-oligonucleotide”, thereby amplifying the di-oligonucleotide, arealso known (Wu et al., Genomics 4:560-569 (1989), the entirety of whichis herein incorporated by reference) and may be readily adapted to thepurposes of the present invention.

Other known nucleic acid amplification procedures, such asallele-specific oligomers, branched DNA technology, transcription-basedamplification systems, or isothermal amplification methods may also beused to amplify and analyze such polymorphisms (Malek et al., U.S. Pat.No. 5,130,238; Davey et al., European Patent Application 329,822;Schuster et al., U.S. Pat. No. 5,169,766; Miller et al., PCT PatentApplication WO 89/06700; Kwoh et al., Proc. Natl. Acad. Sci. (U.S.A.)86:1173-1177 (1989); Gingeras et al., PCT Patent Application WO88/10315; Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:392-396(1992), all of which are herein incorporated by reference in theirentirety).

The identification of a polymorphism can be determined in a variety ofways. By correlating the presence or absence of it in a plant with thepresence or absence of a phenotype, it is possible to predict thephenotype of that plant. If a polymorphism creates or destroys arestriction endonuclease cleavage site, or if it results in the loss orinsertion of DNA (e.g., a VNTR polymorphism), it will alter the size orprofile of the DNA fragments that are generated by digestion with thatrestriction endonuclease. As such, individuals that possess a variantsequence can be distinguished from those having the original sequence byrestriction fragment analysis. Polymorphisms that can be identified inthis manner are termed “restriction fragment length polymorphisms”(“RFLPs”). RFLPs have been widely used in human and plant geneticanalyses (Glassberg, UK Patent Application 2135774; Skolnick et al.,Cytogen. Cell Genet. 32:58-67 (1982); Botstein et al., Ann. J. Hum.Genet. 32:314-331 (1980); Fischer et al., (PCT Application WO90/13668);Uhlen, PCT Application WO90/11369).

Polymorphisms can also be identified by Single Strand ConformationPolymorphism (SSCP) analysis. SSCP is a method capable of identifyingmost sequence variations in a single strand of DNA, typically between150 and 250 nucleotides in length (Elles, Methods in Molecular MedicineMolecular Diagnosis of Genetic Diseases, Humana Press (1996), theentirety of which is herein incorporated by reference); Orita et al.,Genomics 5:874-879 (1989), the entirety of which is herein incorporatedby reference). Under denaturing conditions a single strand of DNA willadopt a conformation that is uniquely dependent on its sequenceconformation. This conformation usually will be different, even if onlya single base is changed. Most conformations have been reported to alterthe physical configuration or size sufficiently to be detectable byelectrophoresis. A number of protocols have been described for SSCPincluding, but not limited to, Lee et al., Anal. Biochem. 205:289-293(1992), the entirety of which is herein incorporated by reference;Suzuki et al., Anal. Biochem. 192:82-84 (1991), the entirety of which isherein incorporated by reference; Lo et al., Nucleic Acids Research20:1005-1009 (1992), the entirety of which is herein incorporated byreference; Sarkar et al., Genomics 13:441-443 (1992), the entirety ofwhich is herein incorporated by reference. It is understood that one ormore of the nucleic acids of the present invention, may be utilized asmarkers or probes to detect polymorphisms by SSCP analysis.

Polymorphisms may also be found using a DNA fingerprinting techniquecalled amplified fragment length polymorphism (AFLP), which is based onthe selective PCR amplification of restriction fragments from a totaldigest of genomic DNA to profile that DNA (Vos et al., Nucleic AcidsRes. 23:4407-4414 (1995), the entirety of which is herein incorporatedby reference). This method allows for the specific co-amplification ofhigh numbers of restriction fragments, which can be visualized by PCRwithout knowledge of the nucleic acid sequence.

AFLP employs basically three steps. Initially, a sample of genomic DNAis cut with restriction enzymes and oligonucleotide adapters are ligatedto the restriction fragments of the DNA. The restriction fragments arethen amplified using PCR by using the adapter and restriction sequenceas target sites for primer annealing. The selective amplification isachieved by the use of primers that extend into the restrictionfragments, amplifying only those fragments in which the primerextensions match the nucleotide flanking the restriction sites. Theseamplified fragments are then visualized on a denaturing polyacrylamidegel.

AFLP analysis has been performed on Salix (Beismann et al., Mol. Ecol.6:989-993 (1997), the entirety of which is herein incorporated byreference), Acinetobacter (Janssen et al., Int. J. Syst. Bacteriol.47:1179-1187 (1997), the entirety of which is herein incorporated byreference), Aeromonas popoffi (Huys et al., Int. J. Syst. Bacteriol.47:1165-1171 (1997), the entirety of which is herein incorporated byreference), rice (McCouch et al., Plant Mol. Biol. 35:89-99 (1997), theentirety of which is herein incorporated by reference; Nandi et al.,Mol. Gen. Genet. 255:1-8 (1997), the entirety of which is hereinincorporated by reference; Cho et al., Genome 39:373-378 (1996), theentirety of which is herein incorporated by reference), barley (Hordeumvulgare) (Simons et al., Genomics 44:61-70 (1997), the entirety of whichis herein incorporated by reference; Waugh et al., Mol. Gen. Genet.255:311-321 (1997), the entirety of which is herein incorporated byreference; Qi et al., Mol. Gen. Genet. 254:330-336 (1997), the entiretyof which is herein incorporated by reference; Becker et al., Mol. Gen.Genet. 249:65-73 (1995), the entirety of which is herein incorporated byreference), potato (Van der Voort et al., Mol. Gen. Genet. 255:438-447(1997), the entirety of which is herein incorporated by reference;Meksem et al., Mol. Gen. Genet. 249:74-81 (1995), the entirety of whichis herein incorporated by reference), Phytophthora infestans (Van derLee et al., Fungal Genet. Biol. 21:278-291 (1997), the entirety of whichis herein incorporated by reference), Bacillus anthracis (Keim et al.,J. Bacteriol. 179:818-824 (1997), the entirety of which is hereinincorporated by reference), Astragalus cremnophylax (Travis et al., Mol.Ecol. 5:735-745 (1996), the entirety of which is herein incorporated byreference), Arabidopsis (Cnops et al., Mol. Gen. Genet. 253:32-41(1996), the entirety of which is herein incorporated by reference),Escherichia coli (Lin et al., Nucleic Acids Res. 24:3649-3650 (1996),the entirety of which is herein incorporated by reference), Aeromonas(Huys et al., Int. J. Syst. Bacteriol. 46:572-580 (1996), the entiretyof which is herein incorporated by reference), nematode (Folkertsma etal., Mol. Plant Microbe Interact. 9:47-54 (1996), the entirety of whichis herein incorporated by reference), tomato (Thomas et al., Plant J.8:785-794 (1995), the entirety of which is herein incorporated byreference) and human (Latorra et al., PCR Methods Appl. 3:351-358(1994), the entirety of which is herein incorporated by reference). AFLPanalysis has also been used for fingerprinting mRNA (Money et al.,Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which is hereinincorporated by reference; Bachem et al., Plant J. 9:745-753 (1996), theentirety of which is herein incorporated by reference). It is understoodthat one or more of the nucleic acids of the present invention, may beutilized as markers or probes to detect polymorphisms by AFLP analysisor for fingerprinting RNA.

Polymorphisms may also be found using random amplified polymorphic DNA(RAPD) (Williams et al., Nucl. Acids Res. 18:6531-6535 (1990), theentirety of which is herein incorporated by reference) and cleaveableamplified polymorphic sequences (CAPS) (Lyamichev et al., Science260:778-783 (1993), the entirety of which is herein incorporated byreference). It is understood that one or more of the nucleic acidmolecules of the present invention, may be utilized as markers or probesto detect polymorphisms by RAPD or CAPS analysis.

Through genetic mapping, a fine scale linkage map can be developed usingDNA markers and, then, a genomic DNA library of large-sized fragmentscan be screened with molecular markers linked to the desired trait.Molecular markers are advantageous for agronomic traits that areotherwise difficult to tag, such as resistance to pathogens, insects andnematodes, tolerance to abiotic stress, quality parameters andquantitative traits such as high yield potential.

The essential requirements for marker-assisted selection in a plantbreeding program are: (1) the marker(s) should co-segregate or beclosely linked with the desired trait; (2) an efficient means ofscreening large populations for the molecular marker(s) should beavailable; and (3) the screening technique should have highreproducibility across laboratories and preferably be economical to useand be user-friendly.

The genetic linkage of marker molecules can be established by a genemapping model such as, without limitation, the flanking marker modelreported by Lander and Botstein, Genetics 121:185-199 (1989) and theinterval mapping, based on maximum likelihood methods described byLander and Botstein, Genetics 121:185-199 (1989) and implemented in thesoftware package MAPMAKER/QTL (Lincoln and Lander, Mapping GenesControlling Quantitative Traits Using MAPMAKER/QTL, Whitehead Institutefor Biomedical Research, Massachusetts, (1990). Additional softwareincludes Qgene, Version 2.23 (1996), Department of Plant Breeding andBiometry, 266 Emerson Hall, Cornell University, Ithaca, N.Y., the manualof which is herein incorporated by reference in its entirety). Use ofQgene software is a particularly preferred approach.

A maximum likelihood estimate (MLE) for the presence of a marker iscalculated, together with an MLE assuming no QTL effect, to avoid falsepositives. A logo of an odds ratio (LOD) is then calculated as:LOD=log₁₀ (MLE for the presence of a QTL/MLE given no linked QTL).

The LOD score essentially indicates how much more likely the data are tohave arisen assuming the presence of a QTL than in its absence. The LODthreshold value for avoiding a false positive with a given confidence,say 95%, depends on the number of markers and the length of the genome.Graphs indicating LOD thresholds are set forth in Lander and Botstein,Genetics 121:185-199 (1989) the entirety of which is herein incorporatedby reference and further described by Arús and Moreno-González, PlantBreeding, Hayward et al., (eds.) Chapman & Hall, London, pp. 314-331(1993), the entirety of which is herein incorporated by reference.

Additional models can be used. Many modifications and alternativeapproaches to interval mapping have been reported, including the usenon-parametric methods (Kruglyak and Lander, Genetics 139:1421-1428(1995), the entirety of which is herein incorporated by reference).Multiple regression methods or models can be also be used, in which thetrait is regressed on a large number of markers (Jansen, Biometrics inPlant Breeding, van Oijen and Jansen (eds.), Proceedings of the NinthMeeting of the Eucarpia Section Biometrics in Plant Breeding, TheNetherlands, pp. 116-124 (1994); Weber and Wricke, Advances in PlantBreeding, Blackwell, Berlin, 16 (1994), both of which is hereinincorporated by reference in their entirety). Procedures combininginterval mapping with regression analysis, whereby the phenotype isregressed onto a single putative QTL at a given marker interval and atthe same time onto a number of markers that serve as ‘cofactors,’ havebeen reported by Jansen and Stam, Genetics 136:1447-1455 (1994), theentirety of which is herein incorporated by reference and Zeng, Genetics136:1457-1468 (1994) the entirety of which is herein incorporated byreference. Generally, the use of cofactors reduces the bias and samplingerror of the estimated QTL positions (Utz and Melchinger, Biometrics inPlant Breeding, van Oijen and Jansen (eds.) Proceedings of the NinthMeeting of the Eucarpia Section Biometrics in Plant Breeding, TheNetherlands, pp. 195-204 (1994), the entirety of which is hereinincorporated by reference, thereby improving the precision andefficiency of QTL mapping (Zeng, Genetics 136:1457-1468 (1994)). Thesemodels can be extended to multi-environment experiments to analyzegenotype-environment interactions (Jansen et al., Theo. Appl. Genet.91:33-37 (1995), the entirety of which is herein incorporated byreference).

Selection of an appropriate mapping populations is important to mapconstruction. The choice of appropriate mapping population depends onthe type of marker systems employed (Tanksley et al., Molecular mappingplant chromosomes. Chromosome structure and function: Impact of newconcepts, Gustafson and Appels (eds.), Plenum Press, New York, pp157-173 (1988), the entirety of which is herein incorporated byreference). Consideration must be given to the source of parents(adapted vs. exotic) used in the mapping population. Chromosome pairingand recombination rates can be severely disturbed (suppressed) in widecrosses (adapted×exotic) and generally yield greatly reduced linkagedistances. Wide crosses will usually provide segregating populationswith a relatively large array of polymorphisms when compared to progenyin a narrow cross (adapted×adapted).

An F₂ population is the first generation of selfing after the hybridseed is produced. Usually a single F₁ plant is selfed to generate apopulation segregating for all the genes in Mendelian (1:2:1) fashion.Maximum genetic information is obtained from a completely classified F₂population using a codominant marker system (Mather, Measurement ofLinkage in Heredity, Methuen and Co., (1938), the entirety of which isherein incorporated by reference). In the case of dominant markers,progeny tests (e.g. F₃, BCF₂) are required to identify theheterozygotes, thus making it equivalent to a completely classified F₂population. However, this procedure is often prohibitive because of thecost and time involved in progeny testing. Progeny testing of F₂individuals is often used in map construction where phenotypes do notconsistently reflect genotype (e.g. disease resistance) or where traitexpression is controlled by a QTL. Segregation data from progeny testpopulations (e.g. F₃ or BCF₂) can be used in map construction.Marker-assisted selection can then be applied to cross progeny based onmarker-trait map associations (F₂, F₃), where linkage groups have notbeen completely disassociated by recombination events (i.e., maximumdisequillibrium).

Recombinant inbred lines (RIL) (genetically related lines; usually >F₅,developed from continuously selfing F₂ lines towards homozygosity) canbe used as a mapping population. Information obtained from dominantmarkers can be maximized by using RIL because all loci are homozygous ornearly so. Under conditions of tight linkage (i.e., about <10%recombination), dominant and co-dominant markers evaluated in RILpopulations provide more information per individual than either markertype in backcross populations (Reiter et al., Proc. Natl. Acad. Sci.(U.S.A.) 89:1477-1481 (1992), the entirety of which is hereinincorporated by reference). However, as the distance between markersbecomes larger (i.e., loci become more independent), the information inRIL populations decreases dramatically when compared to codominantmarkers.

Backcross populations (e.g., generated from a cross between a successfulvariety (recurrent parent) and another variety (donor parent) carrying atrait not present in the former) can be utilized as a mappingpopulation. A series of backcrosses to the recurrent parent can be madeto recover most of its desirable traits. Thus a population is createdconsisting of individuals nearly like the recurrent parent but eachindividual carries varying amounts or mosaic of genomic regions from thedonor parent. Backcross populations can be useful for mapping dominantmarkers if all loci in the recurrent parent are homozygous and the donorand recurrent parent have contrasting polymorphic marker alleles (Reiteret al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)).Information obtained from backcross populations using either codominantor dominant markers is less than that obtained from F₂ populationsbecause one, rather than two, recombinant gametes are sampled per plant.Backcross populations, however, are more informative (at low markersaturation) when compared to RILs as the distance between linked lociincreases in RIL populations (i.e. about 15% recombination). Increasedrecombination can be beneficial for resolution of tight linkages, butmay be undesirable in the construction of maps with low markersaturation.

Near-isogenic lines (NIL) created by many backcrosses to produce anarray of individuals that are nearly identical in genetic compositionexcept for the trait or genomic region under interrogation can be usedas a mapping population. In mapping with NILs, only a portion of thepolymorphic loci are expected to map to a selected region.

Bulk segregant analysis (BSA) is a method developed for the rapididentification of linkage between markers and traits of interest(Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9828-9832 (1991),the entirety of which is herein incorporated by reference). In BSA, twobulked DNA samples are drawn from a segregating population originatingfrom a single cross. These bulks contain individuals that are identicalfor a particular trait (resistant or susceptible to particular disease)or genomic region but arbitrary at unlinked regions (i.e. heterozygous).Regions unlinked to the target region will not differ between the bulkedsamples of many individuals in BSA.

It is understood that one or more of the nucleic acid molecules of thepresent invention may be used as molecular markers. It is alsounderstood that one or more of the protein molecules of the presentinvention may be used as molecular markers.

In accordance with this aspect of the present invention, a samplenucleic acid is obtained from plants cells or tissues. Any source ofnucleic acid may be used. Preferably, the nucleic acid is genomic DNA.The nucleic acid is subjected to restriction endonuclease digestion. Forexample, one or more nucleic acid molecule or fragment thereof of thepresent invention can be used as a probe in accordance with theabove-described polymorphic methods. The polymorphism obtained in thisapproach can then be cloned to identify the mutation at the codingregion which alters the protein's structure or regulatory region of thegene which affects its expression level.

In an aspect of the present invention, one or more of the nucleicmolecules of the present invention are used to determine the level(i.e., the concentration of mRNA in a sample, etc.) in a plant(preferably maize or soybean) or pattern (i.e., the kinetics ofexpression, rate of decomposition, stability profile, etc.) of theexpression of a protein encoded in part or whole by one or more of thenucleic acid molecule of the present invention (collectively, the“Expression Response” of a cell or tissue). As used herein, theExpression Response manifested by a cell or tissue is said to be“altered” if it differs from the Expression Response of cells or tissuesof plants not exhibiting the phenotype. To determine whether aExpression Response is altered, the Expression Response manifested bythe cell or tissue of the plant exhibiting the phenotype is comparedwith that of a similar cell or tissue sample of a plant not exhibitingthe phenotype. As will be appreciated, it is not necessary tore-determine the Expression Response of the cell or tissue sample ofplants not exhibiting the phenotype each time such a comparison is made;rather, the Expression Response of a particular plant may be comparedwith previously obtained values of normal plants. As used herein, thephenotype of the organism is any of one or more characteristics of anorganism (e.g. disease resistance, pest tolerance, environmentaltolerance such as tolerance to abiotic stress, male sterility, qualityimprovement or yield etc.). A change in genotype or phenotype may betransient or permanent. Also as used herein, a tissue sample is anysample that comprises more than one cell. In a preferred aspect, atissue sample comprises cells that share a common characteristic (e.g.derived from root, seed, flower, leaf, stem or pollen etc.).

In one aspect of the present invention, an evaluation can be conductedto determine whether a particular mRNA molecule is present. One or moreof the nucleic acid molecules of the present invention, preferably oneor more of the EST nucleic acid molecules or fragments thereof of thepresent invention are utilized to detect the presence or quantity of themRNA species. Such molecules are then incubated with cell or tissueextracts of a plant under conditions sufficient to permit nucleic acidhybridization. The detection of double-stranded probe-mRNA hybridmolecules is indicative of the presence of the mRNA; the amount of suchhybrid formed is proportional to the amount of mRNA. Thus, such probesmay be used to ascertain the level and extent of the mRNA production ina plant's cells or tissues. Such nucleic acid hybridization may beconducted under quantitative conditions (thereby providing a numericalvalue of the amount of the mRNA present). Alternatively, the assay maybe conducted as a qualitative assay that indicates either that the mRNAis present, or that its level exceeds a user set, predefined value.

A principle of in situ hybridization is that a labeled, single-strandednucleic acid probe will hybridize to a complementary strand of cellularDNA or RNA and, under the appropriate conditions, these molecules willform a stable hybrid. When nucleic acid hybridization is combined withhistological techniques, specific DNA or RNA sequences can be identifiedwithin a single cell. An advantage of in situ hybridization over moreconventional techniques for the detection of nucleic acids is that itallows an investigator to determine the precise spatial population(Angerer et al., Dev. Biol. 101:477-484 (1984), the entirety of which isherein incorporated by reference; Angerer et al., Dev. Biol. 112:157-166(1985), the entirety of which is herein incorporated by reference; Dixonet al., EMBO J. 10:1317-1324 (1991), the entirety of which is hereinincorporated by reference). In situ hybridization may be used to measurethe steady-state level of RNA accumulation. It is a sensitive techniqueand RNA sequences present in as few as 5-10 copies per cell can bedetected (Hardin et al., J. Mol. Biol. 202:417-431 (1989), the entiretyof which is herein incorporated by reference). A number of protocolshave been devised for in situ hybridization, each with tissuepreparation, hybridization and washing conditions (Meyerowitz, PlantMol. Biol. Rep. 5:242-250 (1987), the entirety of which is hereinincorporated by reference; Cox and Goldberg, In: Plant MolecularBiology: A Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford(1988), the entirety of which is herein incorporated by reference;Raikhel et al., In situ RNA hybridization in plant tissues, In: PlantMolecular Biology Manual, vol. B9: 1-32, Kluwer Academic Publisher,Dordrecht, Belgium (1989), the entirety of which is herein incorporatedby reference).

In situ hybridization also allows for the localization of proteinswithin a tissue or cell (Wilkinson, In Situ Hybridization, OxfordUniversity Press, Oxford (1992), the entirety of which is hereinincorporated by reference; Langdale, In Situ Hybridization In: The MaizeHandbook Freeling and Walbot (eds.), pp 165-179, Springer-Verlag, NewYork (1994), the entirety of which is herein incorporated by reference).It is understood that one or more of the molecules of the presentinvention, preferably one or more of the EST nucleic acid molecules orfragments thereof of the present invention or one or more of theantibodies of the present invention may be utilized to detect the levelor pattern of a methionine pathway protein or mRNA thereof by in situhybridization.

Fluorescent in situ hybridization allows the localization of aparticular DNA sequence along a chromosome which is useful, among otheruses, for gene mapping, following chromosomes in hybrid lines ordetecting chromosomes with translocations, transversions or deletions.In situ hybridization has been used to identify chromosomes in severalplant species (Griffor et al., Plant Mol. Biol. 17:101-109 (1991), theentirety of which is herein incorporated by reference; Gustafson et al.,Proc. Natl. Acad. Sci. (U.S.A.) 87:1899-1902 (1990), herein incorporatedby reference; Mukai and Gill, Genome 34:448-452 (1991), the entirety ofwhich is herein incorporated by reference; Schwarzacher andHeslop-Harrison, Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet.66:313-316 (1991), the entirety of which is herein incorporated byreference; Parra and Windle, Nature Genetics 5:17-21 (1993), theentirety of which is herein incorporated by reference). It is understoodthat the nucleic acid molecules of the present invention may be used asprobes or markers to localize sequences along a chromosome.

Another method to localize the expression of a molecule is tissueprinting. Tissue printing provides a way to screen, at the same time onthe same membrane many tissue sections from different plants ordifferent developmental stages. Tissue-printing procedures utilize filmsdesigned to immobilize proteins and nucleic acids. In essence, a freshlycut section of a tissue is pressed gently onto nitrocellulose paper,nylon membrane or polyvinylidene difluoride membrane. Such membranes arecommercially available (e.g. Millipore, Bedford, Mass. U.S.A.). Thecontents of the cut cell transfer onto the membrane and the contents andare immobilized to the membrane. The immobilized contents form a latentprint that can be visualized with appropriate probes. When a planttissue print is made on nitrocellulose paper, the cell walls leave aphysical print that makes the anatomy visible without further treatment(Varner and Taylor, Plant Physiol. 91:31-33 (1989), the entirety ofwhich is herein incorporated by reference).

Tissue printing on substrate films is described by Daoust, Exp. CellRes. 12:203-211 (1957), the entirety of which is herein incorporated byreference, who detected amylase, protease, ribonuclease anddeoxyribonuclease in animal tissues using starch, gelatin and agarfilms. These techniques can be applied to plant tissues (Yomo andTaylor, Planta 112:35-43 (1973); the entirety of which is hereinincorporated by reference; Harris and Chrispeels, Plant Physiol.56:292-299 (1975), the entirety of which is herein incorporated byreference). Advances in membrane technology have increased the range ofapplications of Daoust's tissue-printing techniques allowing (Cassab andVarner, J. Cell. Biol. 105:2581-2588 (1987), the entirety of which isherein incorporated by reference) the histochemical localization ofvarious plant enzymes and deoxyribonuclease on nitrocellulose paper andnylon (Spruce et al., Phytochemistry 26:2901-2903 (1987), the entiretyof which is herein incorporated by reference; Barres et al., Neuron5:527-544 (1990), the entirety of which is herein incorporated byreference; Reid and Pont-Lezica, Tissue Printing: Tools for the Study ofAnatomy, Histochemistry and Gene Expression, Academic Press, New York,N.Y. (1992), the entirety of which is herein incorporated by reference;Reid et al., Plant Physiol. 93:160-165 (1990), the entirety of which isherein incorporated by reference; Ye et al., Plant J. 1:175-183 (1991),the entirety of which is herein incorporated by reference).

It is understood that one or more of the molecules of the presentinvention, preferably one or more of the EST nucleic acid molecules orfragments thereof of the present invention or one or more of theantibodies of the present invention may be utilized to detect thepresence or quantity of a methionine pathway protein by tissue printing.

Further it is also understood that any of the nucleic acid molecules ofthe present invention may be used as marker nucleic acids and or probesin connection with methods that require probes or marker nucleic acids.As used herein, a probe is an agent that is utilized to determine anattribute or feature (e.g. presence or absence, location, correlation,etc.) of a molecule, cell, tissue or plant. As used herein, a markernucleic acid is a nucleic acid molecule that is utilized to determine anattribute or feature (e.g., presence or absence, location, correlation,etc.) or a molecule, cell, tissue or plant.

A microarray-based method for high-throughput monitoring of plant geneexpression may be utilized to measure gene-specific hybridizationtargets. This ‘chip’-based approach involves using microarrays ofnucleic acid molecules as gene-specific hybridization targets toquantitatively measure expression of the corresponding plant genes(Schena et al., Science 270:467-470 (1995), the entirety of which isherein incorporated by reference; Shalon, Ph.D. Thesis, StanfordUniversity (1996), the entirety of which is herein incorporated byreference). Every nucleotide in a large sequence can be queried at thesame time. Hybridization can be used to efficiently analyze nucleotidesequences.

Several microarray methods have been described. One method compares thesequences to be analyzed by hybridization to a set of oligonucleotidesrepresenting all possible subsequences (Bains and Smith, J. Theor. Biol.135:303-307 (1989), the entirety of which is herein incorporated byreference). A second method hybridizes the sample to an array ofoligonucleotide or cDNA molecules. An array consisting ofoligonucleotides complementary to subsequences of a target sequence canbe used to determine the identity of a target sequence, measure itsamount and detect differences between the target and a referencesequence. Nucleic acid molecules microarrays may also be screened withprotein molecules or fragments thereof to determine nucleic acidmolecules that specifically bind protein molecules or fragments thereof.

The microarray approach may be used with polypeptide targets (U.S. Pat.No. 5,445,934; U.S. Pat. No. 5,143,854; U.S. Pat. No. 5,079,600; U.S.Pat. No. 4,923,901, all of which are herein incorporated by reference intheir entirety). Essentially, polypeptides are synthesized on asubstrate (microarray) and these polypeptides can be screened witheither protein molecules or fragments thereof or nucleic acid moleculesin order to screen for either protein molecules or fragments thereof ornucleic acid molecules that specifically bind the target polypeptides.(Fodor et al., Science 251:767-773 (1991), the entirety of which isherein incorporated by reference). It is understood that one or more ofthe nucleic acid molecules or protein or fragments thereof of thepresent invention may be utilized in a microarray based method.

In a preferred embodiment of the present invention microarrays may beprepared that comprise nucleic acid molecules where such nucleic acidmolecules encode at least one, preferably at least two, more preferablyat least three, even more preferably at least four, five six or sevenmethionine pathway enzymes. In a preferred embodiment the nucleic acidmolecules are selected from the group consisting of a nucleic acidmolecule that encodes a maize or a soybean methionineadenosyltransferase enzyme or fragment thereof, a nucleic acid moleculethat encodes a maize or a soybean S-adenosylmethionine decarboxylaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean aspartate kinase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean aspartate-semialdehydedehydrogenase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean O-succinylhomoserine (thiol)-lyase enzymeor fragment thereof, a nucleic acid molecule that encodes a maize or asoybean cystathionine β-lyase enzyme or fragment thereof, a nucleic acidmolecule that encodes a maize or a soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or fragment thereof, a nucleic acid molecule that encodes a maizeor a soybean adenosylhomocysteinase enzyme or fragment thereof, anucleic acid molecule that encodes a maize or a soybean cystathionineβ-synthase enzyme or fragment thereof, a nucleic acid molecule thatencodes a maize or a soybean cystathionine γ-lyase enzyme or fragmentthereof and a nucleic acid molecule that encodes a maize or a soybeanO-acetylhomoserine (thiol)-lyase enzyme or fragment thereof.

Site directed mutagenesis may be utilized to modify nucleic acidsequences, particularly as it is a technique that allows one or more ofthe amino acids encoded by a nucleic acid molecule to be altered (e.g. athreonine to be replaced by a methionine). Three basic methods for sitedirected mutagenesis are often employed. These are cassette mutagenesis(Wells et al., Gene 34:315-323 (1985), the entirety of which is hereinincorporated by reference), primer extension (Gilliam et al., Gene12:129-137 (1980), the entirety of which is herein incorporated byreference; Zoller and Smith, Methods Enzymol. 100:468-500 (1983), theentirety of which is herein incorporated by reference;Dalbadie-McFarland et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413(1982), the entirety of which is herein incorporated by reference) andmethods based upon PCR (Scharf et al., Science 233:1076-1078 (1986), theentirety of which is herein incorporated by reference; Higuchi et al.,Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is hereinincorporated by reference). Site directed mutagenesis approaches arealso described in European Patent 0 385 962, the entirety of which isherein incorporated by reference; European Patent 0 359 472, theentirety of which is herein incorporated by reference; and PCT PatentApplication WO 93/07278, the entirety of which is herein incorporated byreference.

Site directed mutagenesis strategies have been applied to plants forboth in vitro as well as in vivo site directed mutagenesis (Lanz et al.,J. Biol. Chem. 266:9971-9976 (1991), the entirety of which is hereinincorporated by reference; Kovgan and Zhdanov, Biotekhnologiya5:148-154, No. 207160n, Chemical Abstracts 110:225 (1989), the entiretyof which is herein incorporated by reference; Ge et al., Proc. Natl.Acad. Sci. (U.S.A.) 86:4037-4041 (1989), the entirety of which is hereinincorporated by reference; Zhu et al., J. Biol. Chem. 271:18494-18498(1996), the entirety of which is herein incorporated by reference; Chuet al., Biochemistry 33:6150-6157 (1994), the entirety of which isherein incorporated by reference; Small et al., EMBO J. 11:1291-1296(1992), the entirety of which is herein incorporated by reference; Choet al., Mol. Biotechnol. 8:13-16 (1997), the entirety of which is hereinincorporated by reference; Kita et al., J. Biol. Chem. 271:26529-26535(1996), the entirety of which is herein incorporated by reference, Jinet al., Mol. Microbiol. 7:555-562 (1993), the entirety of which isherein incorporated by reference; Hatfield and Vierstra, J. Biol. Chem.267:14799-14803 (1992), the entirety of which is herein incorporated byreference; Zhao et al., Biochemistry 31:5093-5099 (1992), the entiretyof which is herein incorporated by reference).

Any of the nucleic acid molecules of the present invention may either bemodified by site directed mutagenesis or used as, for example, nucleicacid molecules that are used to target other nucleic acid molecules formodification. It is understood that mutants with more than one alterednucleotide can be constructed using techniques that practitioners arefamiliar with such as isolating restriction fragments and ligating suchfragments into an expression vector (see, for example, Sambrook et al.,Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press(1989)).

Sequence-specific DNA-binding proteins play a role in the regulation oftranscription. The isolation of recombinant cDNAs encoding theseproteins facilitates the biochemical analysis of their structural andfunctional properties. Genes encoding such DNA-binding proteins havebeen isolated using classical genetics (Vollbrecht et al., Nature 350:241-243 (1991), the entirety of which is herein incorporated byreference) and molecular biochemical approaches, including the screeningof recombinant cDNA libraries with antibodies (Landschulz et al., GenesDev. 2:786-800 (1988), the entirety of which is herein incorporated byreference) or DNA probes (Bodner et al., Cell 55:505-518 (1988), theentirety of which is herein incorporated by reference). In addition, anin situ screening procedure has been used and has facilitated theisolation of sequence-specific DNA-binding proteins from various plantspecies (Gilmartin et al., Plant Cell 4:839-849 (1992), the entirety ofwhich is herein incorporated by reference; Schindler et al., EMBO J.11:1261-1273 (1992), the entirety of which is herein incorporated byreference). An in situ screening protocol does not require thepurification of the protein of interest (Vinson et al., Genes Dev.2:801-806 (1988), the entirety of which is herein incorporated byreference; Singh et al., Cell 52:415-423 (1988), the entirety of whichis herein incorporated by reference).

Two steps may be employed to characterize DNA-protein interactions. Thefirst is to identify promoter fragments that interact with DNA-bindingproteins, to titrate binding activity, to determine the specificity ofbinding and to determine whether a given DNA-binding activity caninteract with related DNA sequences (Sambrook et al., Molecular Cloning:A Laboratory Manual, 2^(nd) edition, Cold Spring Harbor LaboratoryPress, Cold Spring Harbor, N.Y. (1989)). Electrophoretic mobility-shiftassay is a widely used assay. The assay provides a rapid and sensitivemethod for detecting DNA-binding proteins based on the observation thatthe mobility of a DNA fragment through a nondenaturing, low-ionicstrength polyacrylamide gel is retarded upon association with aDNA-binding protein (Fried and Crother, Nucleic Acids Res. 9:6505-6525(1981), the entirety of which is herein incorporated by reference). Whenone or more specific binding activities have been identified, the exactsequence of the DNA bound by the protein may be determined. Severalprocedures for characterizing protein/DNA-binding sites are used,including methylation and ethylation interference assays (Maxam andGilbert, Methods Enzymol. 65:499-560 (1980), the entirety of which isherein incorporated by reference; Wissman and Hillen, Methods Enzymol.208:365-379 (1991), the entirety of which is herein incorporated byreference), footprinting techniques employing DNase I (Galas andSchmitz, Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which isherein incorporated by reference), 1,10-phenanthroline-copper ionmethods (Sigman et al., Methods Enzymol. 208:414-433 (1991), theentirety of which is herein incorporated by reference) and hydroxylradicals methods (Dixon et al., Methods Enzymol. 208:414-433 (1991), theentirety of which is herein incorporated by reference). It is understoodthat one or more of the nucleic acid molecules of the present inventionmay be utilized to identify a protein or fragment thereof thatspecifically binds to a nucleic acid molecule of the present invention.It is also understood that one or more of the protein molecules orfragments thereof of the present invention may be utilized to identify anucleic acid molecule that specifically binds to it.

A two-hybrid system is based on the fact that many cellular functionsare carried out by proteins, such as transcription factors, thatinteract (physically) with one another. Two-hybrid systems have beenused to probe the function of new proteins (Chien et al., Proc. Natl.Acad. Sci. (U.S.A.) 88:9578-9582 (1991) the entirety of which is hereinincorporated by reference; Durfee et al., Genes Dev. 7:555-569 (1993)the entirety of which is herein incorporated by reference; Choi et al.,Cell 78:499-512 (1994), the entirety of which is herein incorporated byreference; Kranz et al., Genes Dev. 8:313-327 (1994), the entirety ofwhich is herein incorporated by reference).

Interaction mating techniques have facilitated a number of two-hybridstudies of protein-protein interaction. Interaction mating has been usedto examine interactions between small sets of tens of proteins (Finleyand Brent, Proc. Natl. Acad. Sci. (U.S.A.) 91:12098-12984 (1994), theentirety of which is herein incorporated by reference), larger sets ofhundreds of proteins (Bendixen et al., Nucl. Acids Res. 22:1778-1779(1994), the entirety of which is herein incorporated by reference) andto comprehensively map proteins encoded by a small genome (Bartel etal., Nature Genetics 12:72-77 (1996), the entirety of which is hereinincorporated by reference). This technique utilizes proteins fused tothe DNA-binding domain and proteins fused to the activation domain. Theyare expressed in two different haploid yeast strains of opposite matingtype and the strains are mated to determine if the two proteinsinteract. Mating occurs when haploid yeast strains come into contact andresult in the fusion of the two haploids into a diploid yeast strain. Aninteraction can be determined by the activation of a two-hybrid reportergene in the diploid strain. An advantage of this technique is that itreduces the number of yeast transformations needed to test individualinteractions. It is understood that the protein-protein interactions ofprotein or fragments thereof of the present invention may beinvestigated using the two-hybrid system and that any of the nucleicacid molecules of the present invention that encode such proteins orfragments thereof may be used to transform yeast in the two-hybridsystem.

(a) Plant Constructs and Plant Transformants

One or more of the nucleic acid molecules of the present invention maybe used in plant transformation or transfection. Exogenous geneticmaterial may be transferred into a plant cell and the plant cellregenerated into a whole, fertile or sterile plant. Exogenous geneticmaterial is any genetic material, whether naturally occurring orotherwise, from any source that is capable of being inserted into anyorganism. Such genetic material may be transferred into eithermonocotyledons and dicotyledons including, but not limited to maize (pp63-69), soybean (pp 50-60), Arabidopsis (p 45), phaseolus (pp 47-49),peanut (pp 49-50), alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79),oat (pp 80-81), sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p85), fescue (p 85), perennial ryegrass (p 86), sugarcane (p 87),cranberry (p 110), papaya (pp 101-102), banana (p 103), banana (p 103),muskmelon (p 104), apple (p 104), cucumber (p 105), dendrobium (p 109),gladiolus (p 110), chrysanthemum (p 110), liliacea (p 111), cotton (pp113-114), eucalyptus (p 115), sunflower (p 118), canola (p 118),turfgrass (p 121), sugarbeet (p 122), coffee (p 122) and dioscorea (p122), (Christou, In: Particle Bombardment for Genetic Engineering ofPlants, Biotechnology Intelligence Unit. Academic Press, San Diego,Calif. (1996), the entirety of which is herein incorporated byreference).

Transfer of a nucleic acid that encodes for a protein can result inoverexpression of that protein in a transformed cell or transgenicplant. One or more of the proteins or fragments thereof encoded bynucleic acid molecules of the present invention may be overexpressed ina transformed cell or transformed plant. Particularly, any of themethionine pathway proteins or fragments thereof may be overexpressed ina transformed cell or transgenic plant. Such overexpression may be theresult of transient or stable transfer of the exogenous geneticmaterial.

Exogenous genetic material may be transferred into a plant cell and theplant cell by the use of a DNA vector or construct designed for such apurpose. Design of such a vector is generally within the skill of theart (See, Plant Molecular Biology: A Laboratory Manual, Clark (ed.),Springier, New York (1997), the entirety of which is herein incorporatedby reference).

A construct or vector may include a plant promoter to express theprotein or protein fragment of choice. A number of promoters which areactive in plant cells have been described in the literature. Theseinclude the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl.Acad. Sci. (U.S.A.) 84:5745-5749 (1987), the entirety of which is hereinincorporated by reference), the octopine synthase (OCS) promoter (whichare carried on tumor-inducing plasmids of Agrobacterium tumefaciens),the caulimovirus promoters such as the cauliflower mosaic virus (CaMV)19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), theentirety of which is herein incorporated by reference) and the CAMV 35Spromoter (Odell et al., Nature 313:810-812 (1985), the entirety of whichis herein incorporated by reference), the figwort mosaic virus35S-promoter, the light-inducible promoter from the small subunit ofribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter(Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), theentirety of which is herein incorporated by reference), the sucrosesynthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.)87:4144-4148 (1990), the entirety of which is herein incorporated byreference), the R gene complex promoter (Chandler et al., The Plant Cell1:1175-1183 (1989), the entirety of which is herein incorporated byreference) and the chlorophyll a/b binding protein gene promoter, etc.These promoters have been used to create DNA constructs which have beenexpressed in plants; see, e.g., PCT publication WO 84/02913, hereinincorporated by reference in its entirety.

Promoters which are known or are found to cause transcription of DNA inplant cells can be used in the present invention. Such promoters may beobtained from a variety of sources such as plants and plant viruses. Itis preferred that the particular promoter selected should be capable ofcausing sufficient expression to result in the production of aneffective amount of the methionine pathway protein to cause the desiredphenotype. In addition to promoters that are known to causetranscription of DNA in plant cells, other promoters may be identifiedfor use in the current invention by screening a plant cDNA library forgenes which are selectively or preferably expressed in the targettissues or cells.

For the purpose of expression in source tissues of the plant, such asthe leaf, seed, root or stem, it is preferred that the promotersutilized in the present invention have relatively high expression inthese specific tissues. For this purpose, one may choose from a numberof promoters for genes with tissue- or cell-specific or -enhancedexpression. Examples of such promoters reported in the literatureinclude the chloroplast glutamine synthetase GS2 promoter from pea(Edwards et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:3459-3463 (1990),herein incorporated by reference in its entirety), the chloroplastfructose-1,6-biphosphatase (FBPase) promoter from wheat (Lloyd et al.,Mol. Gen. Genet. 225:209-216 (1991), herein incorporated by reference inits entirety), the nuclear photosynthetic ST-LS1 promoter from potato(Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated byreference in its entirety), the serine/threonine kinase (PAL) promoterand the glucoamylase (CHS) promoter from Arabidopsis thaliana. Alsoreported to be active in photosynthetically active tissues are theribulose-1,5-bisphosphate carboxylase (RbcS) promoter from eastern larch(Larix laricina), the promoter for the cab gene, cab6, from pine(Yamamoto et al., Plant Cell Physiol. 35:773-778 (1994), hereinincorporated by reference in its entirety), the promoter for the Cab-1gene from wheat (Fejes et al., Plant Mol. Biol. 15:921-932 (1990),herein incorporated by reference in its entirety), the promoter for theCAB-1 gene from spinach (Lubberstedt et al., Plant Physiol. 104:997-1006(1994), herein incorporated by reference in its entirety), the promoterfor the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981 (1992),the entirety of which is herein incorporated by reference), thepyruvate, orthophosphate dikinase (PPDK) promoter from maize (Matsuokaet al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590 (1993), hereinincorporated by reference in its entirety), the promoter for the tobaccoLhcb1*2 gene (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997), hereinincorporated by reference in its entirety), the Arabidopsis thalianaSUC2 sucrose-H+ symporter promoter (Truernit et al., Planta. 196:564-570(1995), herein incorporated by reference in its entirety) and thepromoter for the thylakoid membrane proteins from spinach (psaD, psaF,psaE, PC, FNR, atpC, atpD, cab, rbcS). Other promoters for thechlorophyll a/b-binding proteins may also be utilized in the presentinvention, such as the promoters for LhcB gene and PsbP gene from whitemustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229(1995), the entirety of which is herein incorporated by reference).

For the purpose of expression in sink tissues of the plant, such as thetuber of the potato plant, the fruit of tomato, or the seed of maize,wheat, rice and barley, it is preferred that the promoters utilized inthe present invention have relatively high expression in these specifictissues. A number of promoters for genes with tuber-specific or-enhanced expression are known, including the class I patatin promoter(Bevan et al., EMBO J. 8:1899-1906 (1986); Jefferson et al., Plant Mol.Biol. 14:995-1006 (1990), both of which are herein incorporated byreference in its entirety), the promoter for the potato tuber ADPGPPgenes, both the large and small subunits, the sucrose synthase promoter(Salanoubat and Belliard, Gene. 60:47-56 (1987), Salanoubat andBelliard, Gene. 84:181-185 (1989), both of which are incorporated byreference in their entirety), the promoter for the major tuber proteinsincluding the 22 kd protein complexes and proteinase inhibitors(Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated byreference in its entirety), the promoter for the granule bound starchsynthase gene (GBSS) (Visser et al., Plant Mol. Biol. 17:691-699 (1991),herein incorporated by reference in its entirety) and other class I andII patatins promoters (Koster-Topfer et al., Mol Gen Genet. 219:390-396(1989); Mignery et al., Gene. 62:27-44 (1988), both of which are hereinincorporated by reference in their entirety).

Other promoters can also be used to express a methionine pathway proteinor fragment thereof in specific tissues, such as seeds or fruits. Thepromoter for β-conglycinin (Chen et al., Dev. Genet. 10: 112-122 (1989),herein incorporated by reference in its entirety) or other seed-specificpromoters such as the napin and phaseolin promoters, can be used. Thezeins are a group of storage proteins found in maize endosperm. Genomicclones for zein genes have been isolated (Pedersen et al., Cell29:1015-1026 (1982), herein incorporated by reference in its entirety)and the promoters from these clones, including the 15 kD, 16 kD, 19 kD,22 kD, 27 kD and γ genes, could also be used. Other promoters known tofunction, for example, in maize include the promoters for the followinggenes: waxy, Brittle, Shrunken 2, Branching enzymes I and II, starchsynthases, debranching enzymes, oleosins, glutelins and sucrosesynthases. A particularly preferred promoter for maize endospermexpression is the promoter for the glutelin gene from rice, moreparticularly the Osgt-1 promoter (Zheng et al., Mol. Cell. Biol.13:5829-5842 (1993), herein incorporated by reference in its entirety).Examples of promoters suitable for expression in wheat include thosepromoters for the ADPglucose pyrosynthase (ADPGPP) subunits, the granulebound and other starch synthase, the branching and debranching enzymes,the embryogenesis-abundant proteins, the gliadins and the glutenins.Examples of such promoters in rice include those promoters for theADPGPP subunits, the granule bound and other starch synthase, thebranching enzymes, the debranching enzymes, sucrose synthases and theglutelins. A particularly preferred promoter is the promoter for riceglutelin, Osgt-1. Examples of such promoters for barley include thosefor the ADPGPP subunits, the granule bound and other starch synthase,the branching enzymes, the debranching enzymes, sucrose synthases, thehordeins, the embryo globulins and the aleurone specific proteins.

Root specific promoters may also be used. An example of such a promoteris the promoter for the acid chitinase gene (Samac et al., Plant Mol.Biol. 25:587-596 (1994), the entirety of which is herein incorporated byreference). Expression in root tissue could also be accomplished byutilizing the root specific subdomains of the CaMV35S promoter that havebeen identified (Lam et al., Proc. Natl. Acad. Sci. (U.S.A.)86:7890-7894 (1989), herein incorporated by reference in its entirety).Other root cell specific promoters include those reported by Conkling etal. (Conkling et al., Plant Physiol. 93:1203-1211 (1990), the entiretyof which is herein incorporated by reference).

Additional promoters that may be utilized are described, for example, inU.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147; 5,447,858; 5,608,144;5,608,144; 5,614,399; 5,633,441; 5,633,435; and 4,633,436, all of whichare herein incorporated in their entirety. In addition, a tissuespecific enhancer may be used (Fromm et al., The Plant Cell 1:977-984(1989), the entirety of which is herein incorporated by reference).

Constructs or vectors may also include with the coding region ofinterest a nucleic acid sequence that acts, in whole or in part, toterminate transcription of that region. For example, such sequences havebeen isolated including the Tr7 3′ sequence and the NOS 3′ sequence(Ingelbrecht et al., The Plant Cell 1:671-680 (1989), the entirety ofwhich is herein incorporated by reference; Bevan et al., Nucleic AcidsRes. 11:369-385 (1983), the entirety of which is herein incorporated byreference), or the like.

A vector or construct may also include regulatory elements. Examples ofsuch include the Adh intron 1 (Callis et al., Genes and Develop.1:1183-1200 (1987), the entirety of which is herein incorporated byreference), the sucrose synthase intron (Vasil et al., Plant Physiol.91:1575-1579 (1989), the entirety of which is herein incorporated byreference) and the TMV omega element (Gallie et al., The Plant Cell1:301-311 (1989), the entirety of which is herein incorporated byreference). These and other regulatory elements may be included whenappropriate.

A vector or construct may also include a selectable marker. Selectablemarkers may also be used to select for plants or plant cells thatcontain the exogenous genetic material. Examples of such include, butare not limited to, a neo gene (Potrykus et al., Mol. Gen. Genet.199:183-188 (1985), the entirety of which is herein incorporated byreference) which codes for kanamycin resistance and can be selected forusing kanamycin, G418, etc.; a bar gene which codes for bialaphosresistance; a mutant EPSP synthase gene (Hinchee et al., Bio/Technology6:915-922 (1988), the entirety of which is herein incorporated byreference) which encodes glyphosate resistance; a nitrilase gene whichconfers resistance to bromoxynil (Stalker et al., J. Biol. Chem.263:6310-6314 (1988), the entirety of which is herein incorporated byreference); a mutant acetolactate synthase gene (ALS) which confersimidazolinone or sulphonylurea resistance (European Patent Application154,204 (Sep. 11, 1985), the entirety of which is herein incorporated byreference); and a methotrexate resistant DHFR gene (Thillet et al., J.Biol. Chem. 263:12500-12508 (1988), the entirety of which is hereinincorporated by reference).

A vector or construct may also include a transit peptide. Incorporationof a suitable chloroplast transit peptide may also be employed (EuropeanPatent Application Publication Number 0218571, the entirety of which isherein incorporated by reference). Translational enhancers may also beincorporated as part of the vector DNA. DNA constructs could contain oneor more 5′ non-translated leader sequences which may serve to enhanceexpression of the gene products from the resulting mRNA transcripts.Such sequences may be derived from the promoter selected to express thegene or can be specifically modified to increase translation of themRNA. Such regions may also be obtained from viral RNAs, from suitableeukaryotic genes, or from a synthetic gene sequence. For a review ofoptimizing expression of transgenes, see Koziel et al., Plant Mol. Biol.32:393-405 (1996), the entirety of which is herein incorporated byreference.

A vector or construct may also include a screenable marker. Screenablemarkers may be used to monitor expression. Exemplary screenable markersinclude a β-glucuronidase or uidA gene (GUS) which encodes an enzyme forwhich various chromogenic substrates are known (Jefferson, Plant Mol.Biol, Rep. 5:387-405 (1987), the entirety of which is hereinincorporated by reference; Jefferson et al., EMBO J. 6:3901-3907 (1987),the entirety of which is herein incorporated by reference); an R-locusgene, which encodes a product that regulates the production ofanthocyanin pigments (red color) in plant tissues (Dellaporta et al.,Stadler Symposium 11:263-282 (1988), the entirety of which is hereinincorporated by reference); a lactamase gene (Sutcliffe et al., Proc.Natl. Acad. Sci. (U.S.A.) 75:3737-3741 (1978), the entirety of which isherein incorporated by reference), a gene which encodes an enzyme forwhich various chromogenic substrates are known (e.g., PADAC, achromogenic cephalosporin); a luciferase gene (Ow et al., Science234:856-859 (1986), the entirety of which is herein incorporated byreference); a xylE gene (Zukowsky et al., Proc. Natl. Acad. Sci.(U.S.A.) 80:1101-1105 (1983), the entirety of which is hereinincorporated by reference) which encodes a catechol diozygenase that canconvert chromogenic catechols; an α-amylase gene (Ikatu et al.,Bio/Technol. 8:241-242 (1990), the entirety of which is hereinincorporated by reference); a tyrosinase gene (Katz et al., J. Gen.Microbiol. 129:2703-2714 (1983), the entirety of which is hereinincorporated by reference) which encodes an enzyme capable of oxidizingtyrosine to DOPA and dopaquinone which in turn condenses to melanin; anα-galactosidase, which will turn a chromogenic α-galactose substrate.

Included within the terms “selectable or screenable marker genes” arealso genes which encode a secretable marker whose secretion can bedetected as a means of identifying or selecting for transformed cells.Examples include markers which encode a secretable antigen that can beidentified by antibody interaction, or even secretable enzymes which canbe detected catalytically. Secretable proteins fall into a number ofclasses, including small, diffusible proteins which are detectable,(e.g., by ELISA), small active enzymes which are detectable inextracellular solution (e.g., α-amylase, β-lactamase, phosphinothricintransferase), or proteins which are inserted or trapped in the cell wall(such as proteins which include a leader sequence such as that found inthe expression unit of extension or tobacco PR-S). Other possibleselectable and/or screenable marker genes will be apparent to those ofskill in the art.

There are many methods for introducing transforming nucleic acidmolecules into plant cells. Suitable methods are believed to includevirtually any method by which nucleic acid molecules may be introducedinto a cell, such as by Agrobacterium infection or direct delivery ofnucleic acid molecules such as, for example, by PEG-mediatedtransformation, by electroporation or by acceleration of DNA coatedparticles, etc (Potrykus, Ann. Rev. Plant Physiol. Plant Mol. Biol.42:205-225 (1991), the entirety of which is herein incorporated byreference; Vasil, Plant Mol. Biol. 25:925-937 (1994), the entirety ofwhich is herein incorporated by reference). For example, electroporationhas been used to transform maize protoplasts (Fromm et al., Nature312:791-793 (1986), the entirety of which is herein incorporated byreference).

Other vector systems suitable for introducing transforming DNA into ahost plant cell include but are not limited to binary artificialchromosome (BIBAC) vectors (Hamilton et al., Gene 200:107-116 (1997),the entirety of which is herein incorporated by reference); andtransfection with RNA viral vectors (Della-Cioppa et al., Ann. N.Y.Acad. Sci. (1996), 792 (Engineering Plants for Commercial Products andApplications), 57-61, the entirety of which is herein incorporated byreference). Additional vector systems also include plant selectable YACvectors such as those described in Mullen et al., Molecular Breeding4:449-457 (1988), the entirety of which is herein incorporated byreference).

Technology for introduction of DNA into cells is well known to those ofskill in the art. Four general methods for delivering a gene into cellshave been described: (1) chemical methods (Graham and van der Eb,Virology 54:536-539 (1973), the entirety of which is herein incorporatedby reference); (2) physical methods such as microinjection (Capecchi,Cell 22:479-488 (1980), the entirety of which is herein incorporated byreference), electroporation (Wong and Neumann, Biochem. Biophys. Res.Commun. 107:584-587 (1982); Fromm et al., Proc. Natl. Acad. Sci.(U.S.A.) 82:5824-5828 (1985); U.S. Pat. No. 5,384,253, all of which areherein incorporated in their entirety); and the gene gun (Johnston andTang, Methods Cell Biol. 43:353-365 (1994), the entirety of which isherein incorporated by reference); (3) viral vectors (Clapp, Clin.Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096(1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all ofwhich are herein incorporated in their entirety); and (4)receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther. 3:147-154(1992), Wagner et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:6099-6103(1992), both of which are incorporated by reference in their entirety).

Acceleration methods that may be used include, for example,microprojectile bombardment and the like. One example of a method fordelivering transforming nucleic acid molecules to plant cells ismicroprojectile bombardment. This method has been reviewed by Yang andChristou (eds.), Particle Bombardment Technology for Gene Transfer,Oxford Press, Oxford, England (1994), the entirety of which is hereinincorporated by reference). Non-biological particles (microprojectiles)that may be coated with nucleic acids and delivered into cells by apropelling force. Exemplary particles include those comprised oftungsten, gold, platinum and the like.

A particular advantage of microprojectile bombardment, in addition to itbeing an effective means of reproducibly transforming monocots, is thatneither the isolation of protoplasts (Cristou et al., Plant Physiol.87:671-674 (1988), the entirety of which is herein incorporated byreference) nor the susceptibility of Agrobacterium infection arerequired. An illustrative embodiment of a method for delivering DNA intomaize cells by acceleration is a biolistics α-particle delivery system,which can be used to propel particles coated with DNA through a screen,such as a stainless steel or Nytex screen, onto a filter surface coveredwith corn cells cultured in suspension. Gordon-Kamm et al., describesthe basic procedure for coating tungsten particles with DNA (Gordon-Kammet al., Plant Cell 2:603-618 (1990), the entirety of which is hereinincorporated by reference). The screen disperses the tungsten nucleicacid particles so that they are not delivered to the recipient cells inlarge aggregates. A particle delivery system suitable for use with thepresent invention is the helium acceleration PDS-1000/He gun isavailable from Bio-Rad Laboratories (Bio-Rad, Hercules, Calif.) (Sanfordet al., Technique 3:3-16 (1991), the entirety of which is hereinincorporated by reference).

For the bombardment, cells in suspension may be concentrated on filters.Filters containing the cells to be bombarded are positioned at anappropriate distance below the microprojectile stopping plate. Ifdesired, one or more screens are also positioned between the gun and thecells to be bombarded.

Alternatively, immature embryos or other target cells may be arranged onsolid culture medium. The cells to be bombarded are positioned at anappropriate distance below the microprojectile stopping plate. Ifdesired, one or more screens are also positioned between theacceleration device and the cells to be bombarded. Through the use oftechniques set forth herein one may obtain up to 1000 or more foci ofcells transiently expressing a marker gene. The number of cells in afocus which express the exogenous gene product 48 hours post-bombardmentoften range from one to ten and average one to three.

In bombardment transformation, one may optimize the pre-bombardmentculturing conditions and the bombardment parameters to yield the maximumnumbers of stable transformants. Both the physical and biologicalparameters for bombardment are important in this technology. Physicalfactors are those that involve manipulating the DNA/microprojectileprecipitate or those that affect the flight and velocity of either themacro- or microprojectiles. Biological factors include all stepsinvolved in manipulation of cells before and immediately afterbombardment, the osmotic adjustment of target cells to help alleviatethe trauma associated with bombardment and also the nature of thetransforming DNA, such as linearized DNA or intact supercoiled plasmids.It is believed that pre-bombardment manipulations are especiallyimportant for successful transformation of immature embryos.

In another alternative embodiment, plastids can be stably transformed.Methods disclosed for plastid transformation in higher plants includethe particle gun delivery of DNA containing a selectable marker andtargeting of the DNA to the plastid genome through homologousrecombination (Svab et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8526-8530(1990); Svab and Maliga, Proc. Natl. Acad. Sci. (U.S.A.) 90:913-917(1993); Staub and Maliga, EMBO J. 12:601-606 (1993); U.S. Pat. Nos.5,451,513 and 5,545,818, all of which are herein incorporated byreference in their entirety).

Accordingly, it is contemplated that one may wish to adjust variousaspects of the bombardment parameters in small scale studies to fullyoptimize the conditions. One may particularly wish to adjust physicalparameters such as gap distance, flight distance, tissue distance andhelium pressure. One may also minimize the trauma reduction factors bymodifying conditions which influence the physiological state of therecipient cells and which may therefore influence transformation andintegration efficiencies. For example, the osmotic state, tissuehydration and the subculture stage or cell cycle of the recipient cellsmay be adjusted for optimum transformation. The execution of otherroutine adjustments will be known to those of skill in the art in lightof the present disclosure.

Agrobacterium-mediated transfer is a widely applicable system forintroducing genes into plant cells because the DNA can be introducedinto whole plant tissues, thereby bypassing the need for regeneration ofan intact plant from a protoplast. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art. See, for example the methods described by Fraley etal., Bio/Technology 3:629-635 (1985) and Rogers et al., Methods Enzymol.153:253-277 (1987), both of which are herein incorporated by referencein their entirety. Further, the integration of the Ti-DNA is arelatively precise process resulting in few rearrangements. The regionof DNA to be transferred is defined by the border sequences andintervening DNA is usually inserted into the plant genome as described(Spielmann et al., Mol. Gen. Genet. 205:34 (1986), the entirety of whichis herein incorporated by reference).

Modern Agrobacterium transformation vectors are capable of replicationin E. coli as well as Agrobacterium, allowing for convenientmanipulations as described (Klee et al., In: Plant DNA InfectiousAgents, Hohn and Schell (eds.), Springer-Verlag, New York, pp. 179-203(1985), the entirety of which is herein incorporated by reference.Moreover, technological advances in vectors for Agrobacterium-mediatedgene transfer have improved the arrangement of genes and restrictionsites in the vectors to facilitate construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes and are suitable for present purposes (Rogers et al.,Methods Enzymol. 153:253-277 (1987)). In addition, Agrobacteriumcontaining both armed and disarmed Ti genes can be used for thetransformations. In those plant strains where Agrobacterium-mediatedtransformation is efficient, it is the method of choice because of thefacile and defined nature of the gene transfer.

A transgenic plant formed using Agrobacterium transformation methodstypically contains a single gene on one chromosome. Such transgenicplants can be referred to as being heterozygous for the added gene. Morepreferred is a transgenic plant that is homozygous for the addedstructural gene; i.e., a transgenic plant that contains two added genes,one gene at the same locus on each chromosome of a chromosome pair. Ahomozygous transgenic plant can be obtained by sexually mating (selfing)an independent segregant transgenic plant that contains a single addedgene, germinating some of the seed produced and analyzing the resultingplants produced for the gene of interest.

It is also to be understood that two different transgenic plants canalso be mated to produce offspring that contain two independentlysegregating added, exogenous genes. Selfing of appropriate progeny canproduce plants that are homozygous for both added, exogenous genes thatencode a polypeptide of interest. Back-crossing to a parental plant andout-crossing with a non-transgenic plant are also contemplated, as isvegetative propagation.

Transformation of plant protoplasts can be achieved using methods basedon calcium phosphate precipitation, polyethylene glycol treatment,electroporation and combinations of these treatments (See, for example,Potrykus et al., Mol. Gen. Genet. 205:193-200 (1986); Lorz et al., Mol.Gen. Genet. 199:178 (1985); Fromm et al., Nature 319:791 (1986);Uchimiya et al., Mol. Gen. Genet. 204:204 (1986); Marcotte et al.,Nature 335:454-457 (1988), all of which are herein incorporated byreference in their entirety).

Application of these systems to different plant strains depends upon theability to regenerate that particular plant strain from protoplasts.Illustrative methods for the regeneration of cereals from protoplastsare described (Fujimura et al., Plant Tissue Culture Letters 2:74(1985); Toriyama et al., Theor Appl. Genet. 205:34 (1986); Yamada etal., Plant Cell Rep. 4:85 (1986); Abdullah et al., Biotechnolog 4:1087(1986), all of which are herein incorporated by reference in theirentirety).

To transform plant strains that cannot be successfully regenerated fromprotoplasts, other ways to introduce DNA into intact cells or tissuescan be utilized. For example, regeneration of cereals from immatureembryos or explants can be effected as described (Vasil, Biotechnology6:397 (1988), the entirety of which is herein incorporated byreference). In addition, “particle gun” or high-velocity microprojectiletechnology can be utilized (Vasil et al., Bio/Technology 10:667 (1992),the entirety of which is herein incorporated by reference).

Using the latter technology, DNA is carried through the cell wall andinto the cytoplasm on the surface of small metal particles as described(Klein et al., Nature 328:70 (1987); Klein et al., Proc. Natl. Acad.Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al., Bio/Technology 6:923(1988), all of which are herein incorporated by reference in theirentirety). The metal particles penetrate through several layers of cellsand thus allow the transformation of cells within tissue explants.

Other methods of cell transformation can also be used and include butare not limited to introduction of DNA into plants by direct DNAtransfer into pollen (Zhou et al., Methods Enzymol. 101:433 (1983); Hesset al., Intern Rev. Cytol. 107:367 (1987); Luo et al., Plant Mol. Biol.Reporter 6:165 (1988), all of which are herein incorporated by referencein their entirety), by direct injection of DNA into reproductive organsof a plant (Pena et al., Nature 325:274 (1987), the entirety of which isherein incorporated by reference), or by direct injection of DNA intothe cells of immature embryos followed by the rehydration of desiccatedembryos (Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entiretyof which is herein incorporated by reference).

The regeneration, development and cultivation of plants from singleplant protoplast transformants or from various transformed explants iswell known in the art (Weissbach and Weissbach, In: Methods for PlantMolecular Biology, Academic Press, San Diego, Calif., (1988), theentirety of which is herein incorporated by reference). Thisregeneration and growth process typically includes the steps ofselection of transformed cells, culturing those individualized cellsthrough the usual stages of embryonic development through the rootedplantlet stage. Transgenic embryos and seeds are similarly regenerated.The resulting transgenic rooted shoots are thereafter planted in anappropriate plant growth medium such as soil.

The development or regeneration of plants containing the foreign,exogenous gene that encodes a protein of interest is well known in theart. Preferably, the regenerated plants are self-pollinated to providehomozygous transgenic plants. Otherwise, pollen obtained from theregenerated plants is crossed to seed-grown plants of agronomicallyimportant lines. Conversely, pollen from plants of these important linesis used to pollinate regenerated plants. A transgenic plant of thepresent invention containing a desired polypeptide is cultivated usingmethods well known to one skilled in the art.

There are a variety of methods for the regeneration of plants from planttissue. The particular method of regeneration will depend on thestarting plant tissue and the particular plant species to beregenerated.

Methods for transforming dicots, primarily by use of Agrobacteriumtumefaciens and obtaining transgenic plants have been published forcotton (U.S. Pat. No. 5,004,863; U.S. Pat. No. 5,159,135; U.S. Pat. No.5,518,908, all of which are herein incorporated by reference in theirentirety); soybean (U.S. Pat. No. 5,569,834; U.S. Pat. No. 5,416,011;McCabe et. al., Biotechnology 6:923 (1988); Christou et al., PlantPhysiol. 87:671-674 (1988); all of which are herein incorporated byreference in their entirety); Brassica (U.S. Pat. No. 5,463,174, theentirety of which is herein incorporated by reference); peanut (Cheng etal., Plant Cell Rep. 15:653-657 (1996), McKently et al., Plant Cell Rep.14:699-703 (1995), all of which are herein incorporated by reference intheir entirety); papaya; and pea (Grant et al., Plant Cell Rep.15:254-258 (1995), the entirety of which is herein incorporated byreference).

Transformation of monocotyledons using electroporation, particlebombardment and Agrobacterium have also been reported. Transformationand plant regeneration have been achieved in asparagus (Bytebier et al.,Proc. Natl. Acad. Sci. (U.S.A.) 84:5354 (1987), the entirety of which isherein incorporated by reference); barley (Wan and Lemaux, Plant Physiol104:37 (1994), the entirety of which is herein incorporated byreference); maize (Rhodes et al., Science 240:204 (1988); Gordon-Kamm etal., Plant Cell 2:603-618 (1990); Fromm et al., Bio/Technology 8:833(1990); Koziel et al., Bio/Technology 11:194 (1993); Armstrong et al.,Crop Science 35:550-557 (1995); all of which are herein incorporated byreference in their entirety); oat (Somers et al., Bio/Technology 10:1589(1992), the entirety of which is herein incorporated by reference);orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988), the entiretyof which is herein incorporated by reference); rice (Toriyama et al.,Theor Appl. Genet. 205:34 (1986); Part et al., Plant Mol. Biol.32:1135-1148 (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133-141(1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al.,Plant Cell Rep. 7:379 (1988); Battraw and Hall, Plant Sci. 86:191-202(1992); Christou et al., Bio/Technology 9:957 (1991), all of which areherein incorporated by reference in their entirety); rye (De la Pena etal., Nature 325:274 (1987), the entirety of which is herein incorporatedby reference); sugarcane (Bower and Birch, Plant J. 2:409 (1992), theentirety of which is herein incorporated by reference); tall fescue(Wang et al., Bio/Technology 10:691 (1992), the entirety of which isherein incorporated by reference) and wheat (Vasil et al.,Bio/Technology 10:667 (1992), the entirety of which is hereinincorporated by reference; U.S. Pat. No. 5,631,152, the entirety ofwhich is herein incorporated by reference.)

Assays for gene expression based on the transient expression of clonednucleic acid constructs have been developed by introducing the nucleicacid molecules into plant cells by polyethylene glycol treatment,electroporation, or particle bombardment (Marcotte et al., Nature335:454-457 (1988), the entirety of which is herein incorporated byreference; Marcotte et al., Plant Cell 1:523-532 (1989), the entirety ofwhich is herein incorporated by reference; McCarty et al., Cell66:895-905 (1991), the entirety of which is herein incorporated byreference; Hattori et al., Genes Dev. 6:609-618 (1992), the entirety ofwhich is herein incorporated by reference; Goff et al., EMBO J.9:2517-2522 (1990), the entirety of which is herein incorporated byreference). Transient expression systems may be used to functionallydissect gene constructs (see generally, Mailga et al., Methods in PlantMolecular Biology, Cold Spring Harbor Press (1995)).

Any of the nucleic acid molecules of the present invention may beintroduced into a plant cell in a permanent or transient manner incombination with other genetic elements such as vectors, promoters,enhancers etc. Further, any of the nucleic acid molecules of the presentinvention may be introduced into a plant cell in a manner that allowsfor overexpression of the protein or fragment thereof encoded by thenucleic acid molecule.

Cosuppression is the reduction in expression levels, usually at thelevel of RNA, of a particular endogenous gene or gene family by theexpression of a homologous sense construct that is capable oftranscribing mRNA of the same strandedness as the transcript of theendogenous gene (Napoli et al., Plant Cell 2:279-289 (1990), theentirety of which is herein incorporated by reference; van der Krol etal., Plant Cell 2:291-299 (1990), the entirety of which is hereinincorporated by reference). Cosuppression may result from stabletransformation with a single copy nucleic acid molecule that ishomologous to a nucleic acid sequence found with the cell (Prolls andMeyer, Plant J. 2:465-475 (1992), the entirety of which is hereinincorporated by reference) or with multiple copies of a nucleic acidmolecule that is homologous to a nucleic acid sequence found with thecell (Mittlesten et al., Mol. Gen. Genet. 244:325-330 (1994), theentirety of which is herein incorporated by reference). Genes, eventhough different, linked to homologous promoters may result in thecosuppression of the linked genes (Vaucheret, C. R. Acad. Sci. III316:1471-1483 (1993), the entirety of which is herein incorporated byreference).

This technique has, for example, been applied to generate white flowersfrom red petunia and tomatoes that do not ripen on the vine. Up to 50%of petunia transformants that contained a sense copy of the glucoamylase(CHS) gene produced white flowers or floral sectors; this was as aresult of the post-transcriptional loss of mRNA encoding CHS (Flavell,Proc. Natl. Acad. Sci. (U.S.A.) 91:3490-3496 (1994), the entirety ofwhich is herein incorporated by reference); van Blokland et al., PlantJ. 6:861-877 (1994), the entirety of which is herein incorporated byreference). Cosuppression may require the coordinate transcription ofthe transgene and the endogenous gene and can be reset by adevelopmental control mechanism (Jorgensen, Trends Biotechnol. 8:340-344(1990), the entirety of which is herein incorporated by reference; Meinsand Kunz, In: Gene Inactivation and Homologous Recombination in Plants,Paszkowski (ed.), pp. 335-348, Kluwer Academic, Netherlands (1994), theentirety of which is herein incorporated by reference).

It is understood that one or more of the nucleic acids of the presentinvention may be introduced into a plant cell and transcribed using anappropriate promoter with such transcription resulting in thecosuppression of an endogenous methionine pathway protein.

Antisense approaches are a way of preventing or reducing gene functionby targeting the genetic material (Mol et al., FEBS Lett. 268:427-430(1990), the entirety of which is herein incorporated by reference). Theobjective of the antisense approach is to use a sequence complementaryto the target gene to block its expression and create a mutant cell lineor organism in which the level of a single chosen protein is selectivelyreduced or abolished. Antisense techniques have several advantages overother ‘reverse genetic’ approaches. The site of inactivation and itsdevelopmental effect can be manipulated by the choice of promoter forantisense genes or by the timing of external application ormicroinjection. Antisense can manipulate its specificity by selectingeither unique regions of the target gene or regions where it shareshomology to other related genes (Hiatt et al., In: Genetic Engineering,Setlow (ed.), Vol. 11, New York: Plenum 49-63 (1989), the entirety ofwhich is herein incorporated by reference).

The principle of regulation by antisense RNA is that RNA that iscomplementary to the target mRNA is introduced into cells, resulting inspecific RNA:RNA duplexes being formed by base pairing between theantisense substrate and the target mRNA (Green et al., Annu. Rev.Biochem. 55:569-597 (1986), the entirety of which is herein incorporatedby reference). Under one embodiment, the process involves theintroduction and expression of an antisense gene sequence. Such asequence is one in which part or all of the normal gene sequences areplaced under a promoter in inverted orientation so that the ‘wrong’ orcomplementary strand is transcribed into a noncoding antisense RNA thathybridizes with the target mRNA and interferes with its expression(Takayama and Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990),the entirety of which is herein incorporated by reference). An antisensevector is constructed by standard procedures and introduced into cellsby transformation, transfection, electroporation, microinjection,infection, etc. The type of transformation and choice of vector willdetermine whether expression is transient or stable. The promoter usedfor the antisense gene may influence the level, timing, tissue,specificity, or inducibility of the antisense inhibition.

It is understood that the activity of a methionine pathway protein in aplant cell may be reduced or depressed by growing a transformed plantcell containing a nucleic acid molecule whose non-transcribed strandencodes a methionine pathway protein or fragment thereof.

Antibodies have been expressed in plants (Hiatt et al., Nature 342:76-78(1989), the entirety of which is herein incorporated by reference;Conrad and Fielder, Plant Mol. Biol. 26:1023-1030 (1994), the entiretyof which is herein incorporated by reference). Cytoplamsic expression ofa scFv (single-chain Fv antibodies) has been reported to delay infectionby artichoke mottled crinkle virus. Transgenic plants that expressantibodies directed against endogenous proteins may exhibit aphysiological effect (Philips et al., EMBO J. 16:4489-4496 (1997), theentirety of which is herein incorporated by reference; Marion-Poll,Trends in Plant Science 2:447-448 (1997), the entirety of which isherein incorporated by reference). For example, expressed anti-abscisicantibodies have been reported to result in a general perturbation ofseed development (Philips et al., EMBO J. 16: 4489-4496 (1997)).

Antibodies that are catalytic may also be expressed in plants (abzymes).The principle behind abzymes is that since antibodies may be raisedagainst many molecules, this recognition ability can be directed towardgenerating antibodies that bind transition states to force a chemicalreaction forward (Persidas, Nature Biotechnology 15:1313-1315 (1997),the entirety of which is herein incorporated by reference; Baca et al.,Ann. Rev. Biophys. Biomol. Struct. 26:461-493 (1997), the entirety ofwhich is herein incorporated by reference). The catalytic abilities ofabzymes may be enhanced by site directed mutagenesis. Examples ofabzymes are, for example, set forth in U.S. Pat. No. 5,658,753; U.S.Pat. No. 5,632,990; U.S. Pat. No. 5,631,137; U.S. Pat. No. 5,602,015;U.S. Pat. No. 5,559,538; U.S. Pat. No. 5,576,174; U.S. Pat. No.5,500,358; U.S. Pat. No. 5,318,897; U.S. Pat. No. 5,298,409; U.S. Pat.No. 5,258,289 and U.S. Pat. No. 5,194,585, all of which are hereinincorporated in their entirety.

It is understood that any of the antibodies of the present invention maybe expressed in plants and that such expression can result in aphysiological effect. It is also understood that any of the expressedantibodies may be catalytic.

(b) Fungal Constructs and Fungal Transformants

The present invention also relates to a fungal recombinant vectorcomprising exogenous genetic material. The present invention alsorelates to a fungal cell comprising a fungal recombinant vector. Thepresent invention also relates to methods for obtaining a recombinantfungal host cell comprising introducing into a fungal host cellexogenous genetic material.

Exogenous genetic material may be transferred into a fungal cell. In apreferred embodiment the exogenous genetic material includes a nucleicacid molecule of the present invention having a sequence selected fromthe group consisting of SEQ ID NO: 1 through SEQ ID NO: 3204 orcomplements thereof or fragments of either. The fungal recombinantvector may be any vector which can be conveniently subjected torecombinant DNA procedures. The choice of a vector will typically dependon the compatibility of the vector with the fungal host cell into whichthe vector is to be introduced. The vector may be a linear or a closedcircular plasmid. The vector system may be a single vector or plasmid ortwo or more vectors or plasmids which together contain the total DNA tobe introduced into the genome of the fungal host.

The fungal vector may be an autonomously replicating vector, i.e., avector which exists as an extrachromosomal entity, the replication ofwhich is independent of chromosomal replication, e.g., a plasmid, anextrachromosomal element, a minichromosome, or an artificial chromosome.The vector may contain any means for assuring self-replication.Alternatively, the vector may be one which, when introduced into thefungal cell, is integrated into the genome and replicated together withthe chromosome(s) into which it has been integrated. For integration,the vector may rely on the nucleic acid sequence of the vector forstable integration of the vector into the genome by homologous ornonhomologous recombination. Alternatively, the vector may containadditional nucleic acid sequences for directing integration byhomologous recombination into the genome of the fungal host. Theadditional nucleic acid sequences enable the vector to be integratedinto the host cell genome at a precise location(s) in the chromosome(s).To increase the likelihood of integration at a precise location, thereshould be preferably two nucleic acid sequences which individuallycontain a sufficient number of nucleic acids, preferably 400 bp to 1500bp, more preferably 800 bp to 1000 bp, which are highly homologous withthe corresponding target sequence to enhance the probability ofhomologous recombination. These nucleic acid sequences may be anysequence that is homologous with a target sequence in the genome of thefungal host cell and, furthermore, may be non-encoding or encodingsequences.

For autonomous replication, the vector may further comprise an origin ofreplication enabling the vector to replicate autonomously in the hostcell in question. Examples of origin of replications for use in a yeasthost cell are the 2 micron origin of replication and the combination ofCEN3 and ARS 1. Any origin of replication may be used which iscompatible with the fungal host cell of choice.

The fungal vectors of the present invention preferably contain one ormore selectable markers which permit easy selection of transformedcells. A selectable marker is a gene the product of which provides, forexample biocide or viral resistance, resistance to heavy metals,prototrophy to auxotrophs and the like. The selectable marker may beselected from the group including, but not limited to, amds(acetamidase), argB (ornithine carbamoyltransferase), bar(phosphinothricin acetyltransferase), hygB (hygromycinphosphotransferase), niaD (nitrate reductase), pyrG(orotidine-5′-phosphate decarboxylase) and sC (sulfateadenyltransferase) and trpC (anthranilate synthase). Preferred for usein an Aspergillus cell are the amdS and pyrG markers of Aspergillusnidulans or Aspergillus oryzae and the bar marker of Streptomyceshygroscopicus. Furthermore, selection may be accomplished byco-transformation, e.g., as described in WO 91/17243, the entirety ofwhich is herein incorporated by reference. A nucleic acid sequence ofthe present invention may be operably linked to a suitable promotersequence. The promoter sequence is a nucleic acid sequence which isrecognized by the fungal host cell for expression of the nucleic acidsequence. The promoter sequence contains transcription and translationcontrol sequences which mediate the expression of the protein orfragment thereof.

A promoter may be any nucleic acid sequence which shows transcriptionalactivity in the fungal host cell of choice and may be obtained fromgenes encoding polypeptides either homologous or heterologous to thehost cell. Examples of suitable promoters for directing thetranscription of a nucleic acid construct of the invention in afilamentous fungal host are promoters obtained from the genes encodingAspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase,Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stablealpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase(glaA), Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease,Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulansacetamidase and hybrids thereof. In a yeast host, a useful promoter isthe Saccharomyces cerevisiae enolase (eno-1) promoter. Particularlypreferred promoters are the TAKA amylase, NA2-tpi (a hybrid of thepromoters from the genes encoding Aspergillus niger neutralalpha-amylase and Aspergillus oryzae triose phosphate isomerase) andglaA promoters.

A protein or fragment thereof encoding nucleic acid molecule of thepresent invention may also be operably linked to a terminator sequenceat its 3′ terminus. The terminator sequence may be native to the nucleicacid sequence encoding the protein or fragment thereof or may beobtained from foreign sources. Any terminator which is functional in thefungal host cell of choice may be used in the present invention, butparticularly preferred terminators are obtained from the genes encodingAspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase,Aspergillus nidulans anthranilate synthase, Aspergillus nigeralpha-glucosidase and Saccharomyces cerevisiae enolase.

A protein or fragment thereof encoding nucleic acid molecule of thepresent invention may also be operably linked to a suitable leadersequence. A leader sequence is a nontranslated region of a mRNA which isimportant for translation by the fungal host. The leader sequence isoperably linked to the 5′ terminus of the nucleic acid sequence encodingthe protein or fragment thereof. The leader sequence may be native tothe nucleic acid sequence encoding the protein or fragment thereof ormay be obtained from foreign sources. Any leader sequence which isfunctional in the fungal host cell of choice may be used in the presentinvention, but particularly preferred leaders are obtained from thegenes encoding Aspergillus oryzae TAKA amylase and Aspergillus oryzaetriose phosphate isomerase.

A polyadenylation sequence may also be operably linked to the 3′terminus of the nucleic acid sequence of the present invention. Thepolyadenylation sequence is a sequence which when transcribed isrecognized by the fungal host to add polyadenosine residues totranscribed mRNA. The polyadenylation sequence may be native to thenucleic acid sequence encoding the protein or fragment thereof or may beobtained from foreign sources. Any polyadenylation sequence which isfunctional in the fungal host of choice may be used in the presentinvention, but particularly preferred polyadenylation sequences areobtained from the genes encoding Aspergillus oryzae TAKA amylase,Aspergillus niger glucoamylase, Aspergillus nidulans anthranilatesynthase and Aspergillus niger alpha-glucosidase.

To avoid the necessity of disrupting the cell to obtain the protein orfragment thereof and to minimize the amount of possible degradation ofthe expressed protein or fragment thereof within the cell, it ispreferred that expression of the protein or fragment thereof gives riseto a product secreted outside the cell. To this end, a protein orfragment thereof of the present invention may be linked to a signalpeptide linked to the amino terminus of the protein or fragment thereof.A signal peptide is an amino acid sequence which permits the secretionof the protein or fragment thereof from the fungal host into the culturemedium. The signal peptide may be native to the protein or fragmentthereof of the invention or may be obtained from foreign sources. The 5′end of the coding sequence of the nucleic acid sequence of the presentinvention may inherently contain a signal peptide coding regionnaturally linked in translation reading frame with the segment of thecoding region which encodes the secreted protein or fragment thereof.Alternatively, the 5′ end of the coding sequence may contain a signalpeptide coding region which is foreign to that portion of the codingsequence which encodes the secreted protein or fragment thereof. Theforeign signal peptide may be required where the coding sequence doesnot normally contain a signal peptide coding region. Alternatively, theforeign signal peptide may simply replace the natural signal peptide toobtain enhanced secretion of the desired protein or fragment thereof.The foreign signal peptide coding region may be obtained from aglucoamylase or an amylase gene from an Aspergillus species, a lipase orproteinase gene from Rhizomucor miehei, the gene for the alpha-factorfrom Saccharomyces cerevisiae, or the calf preprochymosin gene. Aneffective signal peptide for fungal host cells is the Aspergillus oryzaeTAKA amylase signal, Aspergillus niger neutral amylase signal, theRhizomucor miehei aspartic proteinase signal, the Humicola lanuginosuscellulase signal, or the Rhizomucor miehei lipase signal. However, anysignal peptide capable of permitting secretion of the protein orfragment thereof in a fungal host of choice may be used in the presentinvention.

A protein or fragment thereof encoding nucleic acid molecule of thepresent invention may also be linked to a propeptide coding region. Apropeptide is an amino acid sequence found at the amino terminus ofaproprotein or proenzyme. Cleavage of the propeptide from the proproteinyields a mature biochemically active protein. The resulting polypeptideis known as a propolypeptide or proenzyme (or a zymogen in some cases).Propolypeptides are generally inactive and can be converted to matureactive polypeptides by catalytic or autocatalytic cleavage of thepropeptide from the propolypeptide or proenzyme. The propeptide codingregion may be native to the protein or fragment thereof or may beobtained from foreign sources. The foreign propeptide coding region maybe obtained from the Saccharomyces cerevisiae alpha-factor gene orMyceliophthora thermophila laccase gene (WO 95/33836, the entirety ofwhich is herein incorporated by reference).

The procedures used to ligate the elements described above to constructthe recombinant expression vector of the present invention are wellknown to one skilled in the art (see, for example, Sambrook et al.,Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor,N.Y., (1989)).

The present invention also relates to recombinant fungal host cellsproduced by the methods of the present invention which areadvantageously used with the recombinant vector of the presentinvention. The cell is preferably transformed with a vector comprising anucleic acid sequence of the invention followed by integration of thevector into the host chromosome. The choice of fungal host cells will toa large extent depend upon the gene encoding the protein or fragmentthereof and its source. The fungal host cell may, for example, be ayeast cell or a filamentous fungal cell.

“Yeast” as used herein includes Ascosporogenous yeast (Endomycetales),Basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti(Blastomycetes). The Ascosporogenous yeasts are divided into thefamilies Spermophthoraceae and Saccharomycetaceae. The latter iscomprised of four subfamilies, Schizosaccharomycoideae (for example,genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae andSaccharomycoideae (for example, genera Pichia, Kluyveromyces andSaccharomyces). The Basidiosporogenous yeasts include the generaLeucosporidim, Rhodosporidium, Sporidiobolus, Filobasidium andFilobasidiella. Yeast belonging to the Fungi Imperfecti are divided intotwo families, Sporobolomycetaceae (for example, genera Sorobolomyces andBullera) and Cryptococcaceae (for example, genus Candida). Since theclassification of yeast may change in the future, for the purposes ofthis invention, yeast shall be defined as described in Biology andActivities of Yeast (Skinner et al., Soc. App. Bacteriol. SymposiumSeries No. 9, (1980), the entirety of which is herein incorporated byreference). The biology of yeast and manipulation of yeast genetics arewell known in the art (see, for example, Biochemistry and Genetics ofYeast, Bacil et al. (ed.), 2nd edition, 1987; The Yeasts, Rose andHarrison (eds.), 2nd ed., (1987); and The Molecular Biology of the YeastSaccharomyces, Strathern et al. (eds.), (1981), all of which are hereinincorporated by reference in their entirety).

“Fungi” as used herein includes the phyla Ascomycota, Basidiomycota,Chytridiomycota and Zygomycota (as defined by Hawksworth et al., In:Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CABInternational, University Press, Cambridge, UK; the entirety of which isherein incorporated by reference) as well as the Oomycota (as cited inHawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi,8th edition, 1995, CAB International, University Press, Cambridge, UK)and all mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby'sDictionary of The Fungi, 8th edition, 1995, CAB International,University Press, Cambridge, UK). Representative groups of Ascomycotainclude, for example, Neurospora, Eupenicillium (=Penicillium),Emericella (=Aspergillus), Eurotiun (=Aspergillus) and the true yeastslisted above. Examples of Basidiomycota include mushrooms, rusts andsmuts. Representative groups of Chytridiomycota include, for example,Allomyces, Blastocladiella, Coelomomyces and aquatic fungi.Representative groups of Oomycota include, for example,Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examplesof mitosporic fungi include Aspergillus, Penicilliun, Candida andAlternaria. Representative groups of Zygomycota include, for example,Rhizopus and Mucor.

“Filamentous fungi” include all filamentous forms of the subdivisionEumycota and Oomycota (as defined by Hawksworth et al., In: Ainsworthand Bisby's Dictionary of The Fungi, 8th edition, 1995, CABInternational, University Press, Cambridge, UK). The filamentous fungiare characterized by a vegetative mycelium composed of chitin,cellulose, glucan, chitosan, mannan and other complex polysaccharides.Vegetative growth is by hyphal elongation and carbon catabolism isobligately aerobic. In contrast, vegetative growth by yeasts such asSaccharomyces cerevisiae is by budding of a unicellular thallus andcarbon catabolism may be fermentative.

In one embodiment, the fungal host cell is a yeast cell. In a preferredembodiment, the yeast host cell is a cell of the species of Candida,Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia and Yarrowia.In a preferred embodiment, the yeast host cell is a Saccharomycescerevisiae cell, a Saccharomyces carlsbergensis, Saccharomycesdiastaticus cell, a Saccharomyces douglasii cell, a Saccharomyceskluyveri cell, a Saccharomyces norbensis cell, or a Saccharomycesoviformis cell. In another preferred embodiment, the yeast host cell isa Kluyveromyces lactis cell. In another preferred embodiment, the yeasthost cell is a Yarrowia lipolytica cell.

In another embodiment, the fungal host cell is a filamentous fungalcell. In a preferred embodiment, the filamentous fungal host cell is acell of the species of, but not limited to, Acremonium, Aspergillus,Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium,Thielavia, Tolypocladium and Trichoderma. In a preferred embodiment, thefilamentous fungal host cell is an Aspergillus cell. In anotherpreferred embodiment, the filamentous fungal host cell is an Acremoniumcell. In another preferred embodiment, the filamentous fungal host cellis a Fusarium cell. In another preferred embodiment, the filamentousfungal host cell is a Humicola cell. In another preferred embodiment,the filamentous fungal host cell is a Myceliophthora cell. In anothereven preferred embodiment, the filamentous fungal host cell is a Mucorcell. In another preferred embodiment, the filamentous fungal host cellis a Neurospora cell. In another preferred embodiment, the filamentousfungal host cell is a Penicillium cell. In another preferred embodiment,the filamentous fungal host cell is a Thielavia cell. In anotherpreferred embodiment, the filamentous fungal host cell is aTolypocladiun cell. In another preferred embodiment, the filamentousfungal host cell is a Trichoderma cell. In a preferred embodiment, thefilamentous fungal host cell is an Aspergillus oryzae cell, anAspergillus niger cell, an Aspergillus foetidus cell, or an Aspergillusjaponicus cell. In another preferred embodiment, the filamentous fungalhost cell is a Fusarium oxysporum cell or a Fusarium graminearum cell.In another preferred embodiment, the filamentous fungal host cell is aHumicola insolens cell or a Humicola lanuginosus cell. In anotherpreferred embodiment, the filamentous fungal host cell is aMyceliophthora thermophila cell. In a most preferred embodiment, thefilamentous fungal host cell is a Mucor miehei cell. In a most preferredembodiment, the filamentous fungal host cell is a Neurospora crassacell. In a most preferred embodiment, the filamentous fungal host cellis a Penicillium purpurogenum cell. In another most preferredembodiment, the filamentous fungal host cell is a Thielavia terrestriscell. In another most preferred embodiment, the Trichoderma cell is aTrichoderma reesei cell, a Trichoderma viride cell, a Trichodermalongibrachiatum cell, a Trichoderma harzianum cell, or a Trichodermakoningii cell. In a preferred embodiment, the fungal host cell isselected from an A. nidulans cell, an A. niger cell, an A. oryzae celland an A. sojae cell. In a further preferred embodiment, the fungal hostcell is an A. nidulans cell.

The recombinant fungal host cells of the present invention may furthercomprise one or more sequences which encode one or more factors that areadvantageous in the expression of the protein or fragment thereof, forexample, an activator (e.g., a trans-acting factor), a chaperone and aprocessing protease. The nucleic acids encoding one or more of thesefactors are preferably not operably linked to the nucleic acid encodingthe protein or fragment thereof. An activator is a protein whichactivates transcription of a nucleic acid sequence encoding apolypeptide (Kudla et al., EMBO 9:1355-1364 (1990); Jarai and Buxton,Current Genetics 26:2238-244 (1994); Verdier, Yeast 6:271-297 (1990),all of which are herein incorporated by reference in their entirety).The nucleic acid sequence encoding an activator may be obtained from thegenes encoding Saccharomyces cerevisiae heme activator protein 1 (hap1),Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4) andAspergillus nidulans ammonia regulation protein (areA). For furtherexamples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et al., Journalof Gen. Microbiol. 139:2295-2307 (1993), both of which are hereinincorporated by reference in their entirety). A chaperone is a proteinwhich assists another protein in folding properly (Hartl et al., TIBS19:20-25 (1994); Bergeron et al., TIBS 19:124-128 (1994); Demolder etal., J. Biotechnology 32:179-189 (1994); Craig, Science 260:1902-1903(1993); Gething and Sambrook, Nature 355:33-45 (1992); Puig and Gilbert,J. Biol. Chem. 269:7764-7771 (1994); Wang and Tsou, FASEB Journal7:1515-11157 (1993); Robinson et al., Bio/Technology 1:381-384 (1994),all of which are herein incorporated by reference in their entirety).The nucleic acid sequence encoding a chaperone may be obtained from thegenes encoding Aspergillus oryzae protein disulphide isomerase,Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae BiP/GRP78and Saccharomyces cerevisiae Hsp70. For further examples, see Gethingand Sambrook, Nature 355:33-45 (1992); Hartl et al., TIBS 19:20-25(1994). A processing protease is a protease that cleaves a propeptide togenerate a mature biochemically active polypeptide (Enderlin andOgrydziak, Yeast 10:67-79 (1994); Fuller et al., Proc. Natl. Acad. Sci.(U.S.A.) 86:1434-1438 (1989); Julius et al., Cell 37:1075-1089 (1984);Julius et al., Cell 32:839-852 (1983), all of which are incorporated byreference in their entirety). The nucleic acid sequence encoding aprocessing protease may be obtained from the genes encoding Aspergillusniger Kex2, Saccharomyces cerevisiae dipeptidylaminopeptidase,Saccharomyces cerevisiae Kex2 and Yarrowia lipolytica dibasic processingendoprotease (xpr6). Any factor that is functional in the fungal hostcell of choice may be used in the present invention.

Fungal cells may be transformed by a process involving protoplastformation, transformation of the protoplasts and regeneration of thecell wall in a manner known per se. Suitable procedures fortransformation of Aspergillus host cells are described in EP 238 023 andYelton et al., Proc. Natl. Acad. Sci. (U.S.A.) 81:1470-1474 (1984), bothof which are herein incorporated by reference in their entirety. Asuitable method of transforming Fusarium species is described byMalardier et al., Gene 78:147-156 (1989), the entirety of which isherein incorporated by reference. Yeast may be transformed using theprocedures described by Becker and Guarente, In: Abelson and Simon,(eds.), Guide to Yeast Genetics and Molecular Biology, Methods Enzymol.Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., J.Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci.(U.S.A.) 75:1920 (1978), all of which are herein incorporated byreference in their entirety.

The present invention also relates to methods of producing the proteinor fragment thereof comprising culturing the recombinant fungal hostcells under conditions conducive for expression of the protein orfragment thereof. The fungal cells of the present invention arecultivated in a nutrient medium suitable for production of the proteinor fragment thereof using methods known in the art. For example, thecell may be cultivated by shake flask cultivation, small-scale orlarge-scale fermentation (including continuous, batch, fed-batch, orsolid state fermentations) in laboratory or industrial fermentorsperformed in a suitable medium and under conditions allowing the proteinor fragment thereof to be expressed and/or isolated. The cultivationtakes place in a suitable nutrient medium comprising carbon and nitrogensources and inorganic salts, using procedures known in the art (see,e.g., Bennett and LaSure (eds.), More Gene Manipulations in Fungi,Academic Press, CA, (1991), the entirety of which is herein incorporatedby reference). Suitable media are available from commercial suppliers ormay be prepared according to published compositions (e.g., in cataloguesof the American Type Culture Collection, Manassas, Va.). If the proteinor fragment thereof is secreted into the nutrient medium, a protein orfragment thereof can be recovered directly from the medium. If theprotein or fragment thereof is not secreted, it is recovered from celllysates.

The expressed protein or fragment thereof may be detected using methodsknown in the art that are specific for the particular protein orfragment. These detection methods may include the use of specificantibodies, formation of an enzyme product, or disappearance of anenzyme substrate. For example, if the protein or fragment thereof hasenzymatic activity, an enzyme assay may be used. Alternatively, ifpolyclonal or monoclonal antibodies specific to the protein or fragmentthereof are available, immunoassays may be employed using the antibodiesto the protein or fragment thereof. The techniques of enzyme assay andimmunoassay are well known to those skilled in the art.

The resulting protein or fragment thereof may be recovered by methodsknown in the arts. For example, the protein or fragment thereof may berecovered from the nutrient medium by conventional procedures including,but not limited to, centrifugation, filtration, extraction,spray-drying, evaporation, or precipitation. The recovered protein orfragment thereof may then be further purified by a variety ofchromatographic procedures, e.g., ion exchange chromatography, gelfiltration chromatography, affinity chromatography, or the like.

(c) Mammalian Constructs and Transformed Mammalian Cells

The present invention also relates to methods for obtaining arecombinant mammalian host cell, comprising introducing into a mammalianhost cell exogenous genetic material. The present invention also relatesto a mammalian cell comprising a mammalian recombinant vector. Thepresent invention also relates to methods for obtaining a recombinantmammalian host cell, comprising introducing into a mammalian cellexogenous genetic material.

Mammalian cell lines available as hosts for expression are known in theart and include many immortalized cell lines available from the AmericanType Culture Collection (ATCC, Manassas, Va.), such as HeLa cells,Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells and anumber of other cell lines. Suitable promoters for mammalian cells arealso known in the art and include viral promoters such as that fromSimian Virus 40 (SV40) (Fiers et al., Nature 273:113 (1978), theentirety of which is herein incorporated by reference), Rous sarcomavirus (RSV), adenovirus (ADV) and bovine papilloma virus (BPV).Mammalian cells may also require terminator sequences and poly-Aaddition sequences. Enhancer sequences which increase expression mayalso be included and sequences which promote amplification of the genemay also be desirable (for example methotrexate resistance genes).

Vectors suitable for replication in mammalian cells may include viralreplicons, or sequences which insure integration of the appropriatesequences encoding HCV epitopes into the host genome. For example,another vector used to express foreign DNA is vaccinia virus. In thiscase, for example, a nucleic acid molecule encoding a protein orfragment thereof is inserted into the vaccinia genome. Techniques forthe insertion of foreign DNA into the vaccinia virus genome are known inthe art and may utilize, for example, homologous recombination. Suchheterologous DNA is generally inserted into a gene which isnon-essential to the virus, for example, the thymidine kinase gene (tk),which also provides a selectable marker. Plasmid vectors that greatlyfacilitate the construction of recombinant viruses have been described(see, for example, Mackett et al., J. Virol. 49:857 (1984); Chakrabartiet al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer VectorsFor Mammalian Cells (Miller and Calos, eds., Cold Spring HarborLaboratory, N.Y., p. 10, (1987); all of which are herein incorporated byreference in their entirety). Expression of the HCV polypeptide thenoccurs in cells or animals which are infected with the live recombinantvaccinia virus.

The sequence to be integrated into the mammalian sequence may beintroduced into the primary host by any convenient means, which includescalcium precipitated DNA, spheroplast fusion, transformation,electroporation, biolistics, lipofection, microinjection, or otherconvenient means. Where an amplifiable gene is being employed, theamplifiable gene may serve as the selection marker for selecting hostsinto which the amplifiable gene has been introduced. Alternatively, onemay include with the amplifiable gene another marker, such as a drugresistance marker, e.g. neomycin resistance (G418 in mammalian cells),hygromycin in resistance etc., or an auxotrophy marker (HIS3, TRP1,LEU2, URA3, ADE2, LYS2, etc.) for use in yeast cells.

Depending upon the nature of the modification and associated targetingconstruct, various techniques may be employed for identifying targetedintegration. Conveniently, the DNA may be digested with one or morerestriction enzymes and the fragments probed with an appropriate DNAfragment which will identify the properly sized restriction fragmentassociated with integration.

One may use different promoter sequences, enhancer sequences, or othersequence which will allow for enhanced levels of expression in theexpression host. Thus, one may combine an enhancer from one source, apromoter region from another source, a 5′-noncoding region upstream fromthe initiation methionine from the same or different source as the othersequences and the like. One may provide for an intron in the non-codingregion with appropriate splice sites or for an alternative3′-untranslated sequence or polyadenylation site. Depending upon theparticular purpose of the modification, any of these sequences may beintroduced, as desired.

Where selection is intended, the sequence to be integrated will havewith it a marker gene, which allows for selection. The marker gene mayconveniently be downstream from the target gene and may includeresistance to a cytotoxic agent, e.g. antibiotics, heavy metals, or thelike, resistance or susceptibility to HAT, gancyclovir, etc.,complementation to an auxotrophic host, particularly by using anauxotrophic yeast as the host for the subject manipulations, or thelike. The marker gene may also be on a separate DNA molecule,particularly with primary mammalian cells. Alternatively, one may screenthe various transformants, due to the high efficiency of recombinationin yeast, by using hybridization analysis, PCR, sequencing, or the like.

For homologous recombination, constructs can be prepared where theamplifiable gene will be flanked, normally on both sides with DNAhomologous with the DNA of the target region. Depending upon the natureof the integrating DNA and the purpose of the integration, thehomologous DNA will generally be within 100 kb, usually 50 kb,preferably about 25 kb, of the transcribed region of the target gene,more preferably within 2 kb of the target gene. Where modeling of thegene is intended, homology will usually be present proximal to the siteof the mutation. The homologous DNA may include the 5′-upstream regionoutside of the transcriptional regulatory region or comprising anyenhancer sequences, transcriptional initiation sequences, adjacentsequences, or the like. The homologous region may include a portion ofthe coding region, where the coding region may be comprised only of anopen reading frame or combination of exons and introns. The homologousregion may comprise all or a portion of an intron, where all or aportion of one or more exons may also be present. Alternatively, thehomologous region may comprise the 3′-region, so as to comprise all or aportion of the transcriptional termination region, or the region 3′ ofthis region. The homologous regions may extend over all or a portion ofthe target gene or be outside the target gene comprising all or aportion of the transcriptional regulatory regions and/or the structuralgene.

The integrating constructs may be prepared in accordance withconventional ways, where sequences may be synthesized, isolated fromnatural sources, manipulated, cloned, ligated, subjected to in vitromutagenesis, primer repair, or the like. At various stages, the joinedsequences may be cloned and analyzed by restriction analysis,sequencing, or the like. Usually during the preparation of a constructwhere various fragments are joined, the fragments, intermediateconstructs and constructs will be carried on a cloning vector comprisinga replication system functional in a prokaryotic host, e.g., E. coli anda marker for selection, e.g., biocide resistance, complementation to anauxotrophic host, etc. Other functional sequences may also be present,such as polylinkers, for ease of introduction and excision of theconstruct or portions thereof, or the like. A large number of cloningvectors are available such as pBR322, the pUC series, etc. Theseconstructs may then be used for integration into the primary mammalianhost.

In the case of the primary mammalian host, a replicating vector may beused. Usually, such vector will have a viral replication system, such asSV40, bovine papilloma virus, adenovirus, or the like. The linear DNAsequence vector may also have a selectable marker for identifyingtransfected cells. Selectable markers include the neo gene, allowing forselection with G418, the herpes tk gene for selection with HAT medium,the gpt gene with mycophenolic acid, complementation of an auxotrophichost, etc.

The vector may or may not be capable of stable maintenance in the host.Where the vector is capable of stable maintenance, the cells will bescreened for homologous integration of the vector into the genome of thehost, where various techniques for curing the cells may be employed.Where the vector is not capable of stable maintenance, for example,where a temperature sensitive replication system is employed, one maychange the temperature from the permissive temperature to thenon-permissive temperature, so that the cells may be cured of thevector. In this case, only those cells having integration of theconstruct comprising the amplifiable gene and, when present, theselectable marker, will be able to survive selection.

Where a selectable marker is present, one may select for the presence ofthe targeting construct by means of the selectable marker. Where theselectable marker is not present, one may select for the presence of theconstruct by the amplifiable gene. For the neo gene or the herpes tkgene, one could employ a medium for growth of the transformants of about0.1-1 mg/ml of G418 or may use HAT medium, respectively. Where DHFR isthe amplifiable gene, the selective medium may include from about0.01-0.5 μM of methotrexate or be deficient inglycine-hypoxanthine-thymidine and have dialysed serum (GHT media).

The DNA can be introduced into the expression host by a variety oftechniques that include calcium phosphate/DNA co-precipitates,microinjection of DNA into the nucleus, electroporation, yeastprotoplast fusion with intact cells, transfection, polycations, e.g.,polybrene, polyornithine, etc., or the like. The DNA may be single ordouble stranded DNA, linear or circular. The various techniques fortransforming mammalian cells are well known (see Keown et al., MethodsEnzymol. (1989); Keown et al., Methods Enzymol. 185:527-537 (1990);Mansour et al., Nature 336:348-352, (1988); all of which are hereinincorporated by reference in their entirety).

(d) Insect Constructs and Transformed Insect Cells

The present invention also relates to an insect recombinant vectorscomprising exogenous genetic material. The present invention alsorelates to an insect cell comprising an insect recombinant vector. Thepresent invention also relates to methods for obtaining a recombinantinsect host cell, comprising introducing into an insect cell exogenousgenetic material.

The insect recombinant vector may be any vector which can beconveniently subjected to recombinant DNA procedures and can bring aboutthe expression of the nucleic acid sequence. The choice of a vector willtypically depend on the compatibility of the vector with the insect hostcell into which the vector is to be introduced. The vector may be alinear or a closed circular plasmid. The vector system may be a singlevector or plasmid or two or more vectors or plasmids which togethercontain the total DNA to be introduced into the genome of the insecthost. In addition, the insect vector may be an expression vector.Nucleic acid molecules can be suitably inserted into a replicationvector for expression in the insect cell under a suitable promoter forinsect cells. Many vectors are available for this purpose and selectionof the appropriate vector will depend mainly on the size of the nucleicacid molecule to be inserted into the vector and the particular hostcell to be transformed with the vector. Each vector contains variouscomponents depending on its function (amplification of DNA or expressionof DNA) and the particular host cell with which it is compatible. Thevector components for insect cell transformation generally include, butare not limited to, one or more of the following: a signal sequence,origin of replication, one or more marker genes and an induciblepromoter.

The insect vector may be an autonomously replicating vector, i.e., avector which exists as an extrachromosomal entity, the replication ofwhich is independent of chromosomal replication, e.g., a plasmid, anextrachromosomal element, a minichromosome, or an artificial chromosome.The vector may contain any means for assuring self-replication.Alternatively, the vector may be one which, when introduced into theinsect cell, is integrated into the genome and replicated together withthe chromosome(s) into which it has been integrated. For integration,the vector may rely on the nucleic acid sequence of the vector forstable integration of the vector into the genome by homologous ornonhomologous recombination. Alternatively, the vector may containadditional nucleic acid sequences for directing integration byhomologous recombination into the genome of the insect host. Theadditional nucleic acid sequences enable the vector to be integratedinto the host cell genome at a precise location(s) in the chromosome(s).To increase the likelihood of integration at a precise location, thereshould be preferably two nucleic acid sequences which individuallycontain a sufficient number of nucleic acids, preferably 400 bp to 1500bp, more preferably 800 bp to 1000 bp, which are highly homologous withthe corresponding target sequence to enhance the probability ofhomologous recombination. These nucleic acid sequences may be anysequence that is homologous with a target sequence in the genome of theinsect host cell and, furthermore, may be non-encoding or encodingsequences.

Baculovirus expression vectors (BEVs) have become important tools forthe expression of foreign genes, both for basic research and for theproduction of proteins with direct clinical applications in human andveterinary medicine (Doerfler, Curr. Top. Microbiol. Immunol. 131:51-68(1968); Luckow and Summers, Bio/Technology 6:47-55 (1988a); Miller,Annual Review of Microbiol. 42:177-199 (1988); Summers, Curr. Comm.Molecular Biology, Cold Spring Harbor Press, Cold Spring Harbor, N.Y.(1988); all of which are herein incorporated by reference in theirentirety). BEVs are recombinant insect viruses in which the codingsequence for a chosen foreign gene has been inserted behind abaculovirus promoter in place of the viral gene, e.g., polyhedrin (Smithand Summers, U.S. Pat. No. 4,745,051, the entirety of which isincorporated herein by reference).

The use of baculovirus vectors relies upon the host cells being derivedfrom Lepidopteran insects such as Spodoptera frugiperda or Trichoplusiani. The preferred Spodoptera frugiperda cell line is the cell line Sf9.The Spodoptera frugiperda Sf9 cell line was obtained from American TypeCulture Collection (Manassas, Va.) and is assigned accession number ATCCCRL 1711 (Summers and Smith, A Manual of Methods for Baculovirus Vectorsand Insect Cell Culture Procedures, Texas Ag. Exper. Station BulletinNo. 1555 (1988), the entirety of which is herein incorporated byreference). Other insect cell systems, such as the silkworm B. mori mayalso be used.

The proteins expressed by the BEVs are, therefore, synthesized, modifiedand transported in host cells derived from Lepidopteran insects. Most ofthe genes that have been inserted and produced in the baculovirusexpression vector system have been derived from vertebrate species.Other baculovirus genes in addition to the polyhedrin promoter may beemployed to advantage in a baculovirus expression system. These includeimmediate-early (alpha), delayed-early (β), late (γ), or very late(delta), according to the phase of the viral infection during which theyare expressed. The expression of these genes occurs sequentially,probably as the result of a “cascade” mechanism of transcriptionalregulation. (Guarino and Summers, J. Virol. 57:563-571 (1986); Guarinoand Summers, J. Virol. 61:2091-2099 (1987); Guarino and Summers, Virol.162:444-451 (1988); all of which are herein incorporated by reference intheir entirety).

Insect recombinant vectors are useful as intermediates for the infectionor transformation of insect cell systems. For example, an insectrecombinant vector containing a nucleic acid molecule encoding abaculovirus transcriptional promoter followed downstream by an insectsignal DNA sequence is capable of directing the secretion of the desiredbiologically active protein from the insect cell. The vector may utilizea baculovirus transcriptional promoter region derived from any of theover 500 baculoviruses generally infecting insects, such as for examplethe Orders Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera,including for example but not limited to the viral DNAs of Autographacalifornica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV, Rachiplusia ouMNPV or Galleria mellonella MNPV, wherein said baculovirustranscriptional promoter is a baculovirus immediate-early gene IEI orIEN promoter; an immediate-early gene in combination with a baculovirusdelayed-early gene promoter region selected from the group consisting of39K and a HindIII-k fragment delayed-early gene; or a baculovirus lategene promoter. The immediate-early or delayed-early promoters can beenhanced with transcriptional enhancer elements. The insect signal DNAsequence may code for a signal peptide of a Lepidopteran adipokinetichormone precursor or a signal peptide of the Manduca sexta adipokinetichormone precursor (Summers, U.S. Pat. No. 5,155,037; the entirety ofwhich is herein incorporated by reference). Other insect signal DNAsequences include a signal peptide of the Orthoptera Schistocercagregaria locust adipokinetic hormone precurser and the Drosophilamelanogaster cuticle genes CP1, CP2, CP3 or CP4 or for an insect signalpeptide having substantially a similar chemical composition and function(Summers, U.S. Pat. No. 5,155,037).

Insect cells are distinctly different from animal cells. Insects have aunique life cycle and have distinct cellular properties such as the lackof intracellular plasminogen activators in which are present invertebrate cells. Another difference is the high expression levels ofprotein products ranging from 1 to greater than 500 mg/liter and theease at which cDNA can be cloned into cells (Frasier, In Vitro Cell.Dev. Biol. 25:225 (1989); Summers and Smith, In: A Manual of Methods forBaculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper.Station Bulletin No. 1555 (1988), both of which are incorporated byreference in their entirety).

Recombinant protein expression in insect cells is achieved by viralinfection or stable transformation. For viral infection, the desiredgene is cloned into baculovirus at the site of the wild-type polyhedrongene (Webb and Summers, Technique 2:173 (1990); Bishop and Posse, Adv.Gene Technol. 1:55 (1990); both of which are incorporated by referencein their entirety). The polyhedron gene is a component of a protein coatin occlusions which encapsulate virus particles. Deletion or insertionin the polyhedron gene results the failure to form occlusion bodies.Occlusion negative viruses are morphologically different from occlusionpositive viruses and enable one skilled in the art to identify andpurify recombinant viruses.

The vectors of present invention preferably contain one or moreselectable markers which permit easy selection of transformed cells. Aselectable marker is a gene the product of which provides, for examplebiocide or viral resistance, resistance to heavy metals, prototrophy toauxotrophs and the like. Selection may be accomplished byco-transformation, e.g., as described in WO 91/17243, a nucleic acidsequence of the present invention may be operably linked to a suitablepromoter sequence. The promoter sequence is a nucleic acid sequencewhich is recognized by the insect host cell for expression of thenucleic acid sequence. The promoter sequence contains transcription andtranslation control sequences which mediate the expression of theprotein or fragment thereof. The promoter may be any nucleic acidsequence which shows transcriptional activity in the insect host cell ofchoice and may be obtained from genes encoding polypeptides eitherhomologous or heterologous to the host cell.

For example, a nucleic acid molecule encoding a protein or fragmentthereof may also be operably linked to a suitable leader sequence. Aleader sequence is a nontranslated region of a mRNA which is importantfor translation by the fungal host. The leader sequence is operablylinked to the 5′ terminus of the nucleic acid sequence encoding theprotein or fragment thereof. The leader sequence may be native to thenucleic acid sequence encoding the protein or fragment thereof or may beobtained from foreign sources. Any leader sequence which is functionalin the insect host cell of choice may be used in the present invention.

A polyadenylation sequence may also be operably linked to the 3′terminus of the nucleic acid sequence of the present invention. Thepolyadenylation sequence is a sequence which when transcribed isrecognized by the insect host to add polyadenosine residues totranscribed mRNA. The polyadenylation sequence may be native to thenucleic acid sequence encoding the protein or fragment thereof or may beobtained from foreign sources. Any polyadenylation sequence which isfunctional in the fungal host of choice may be used in the presentinvention.

To avoid the necessity of disrupting the cell to obtain the protein orfragment thereof and to minimize the amount of possible degradation ofthe expressed polypeptide within the cell, it is preferred thatexpression of the polypeptide gene gives rise to a product secretedoutside the cell. To this end, the protein or fragment thereof of thepresent invention may be linked to a signal peptide linked to the aminoterminus of the protein or fragment thereof. A signal peptide is anamino acid sequence which permits the secretion of the protein orfragment thereof from the insect host into the culture medium. Thesignal peptide may be native to the protein or fragment thereof of theinvention or may be obtained from foreign sources. The 5′ end of thecoding sequence of the nucleic acid sequence of the present inventionmay inherently contain a signal peptide coding region naturally linkedin translation reading frame with the segment of the coding region whichencodes the secreted protein or fragment thereof.

At present, a mode of achieving secretion of a foreign gene product ininsect cells is by way of the foreign gene's native signal peptide.Because the foreign genes are usually from non-insect organisms, theirsignal sequences may be poorly recognized by insect cells and hence,levels of expression may be suboptimal. However, the efficiency ofexpression of foreign gene products seems to depend primarily on thecharacteristics of the foreign protein. On average, nuclear localized ornon-structural proteins are most highly expressed, secreted proteins areintermediate and integral membrane proteins are the least expressed. Onefactor generally affecting the efficiency of the production of foreigngene products in a heterologous host system is the presence of nativesignal sequences (also termed presequences, targeting signals, or leadersequences) associated with the foreign gene. The signal sequence isgenerally coded by a DNA sequence immediately following (5′ to 3′) thetranslation start site of the desired foreign gene.

The expression dependence on the type of signal sequence associated witha gene product can be represented by the following example: If a foreigngene is inserted at a site downstream from the translational start siteof the baculovirus polyhedrin gene so as to produce a fusion protein(containing the N-terminus of the polyhedrin structural gene), the fusedgene is highly expressed. But less expression is achieved when a foreigngene is inserted in a baculovirus expression vector immediatelyfollowing the transcriptional start site and totally replacing thepolyhedrin structural gene.

Insertions into the region −50 to −1 significantly alter (reduce) steadystate transcription which, in turn, reduces translation of the foreigngene product. Use of the pVL941 vector optimizes transcription offoreign genes to the level of the polyhedrin gene transcription. Eventhough the transcription of a foreign gene may be optimal, optimaltranslation may vary because of several factors involving processing:signal peptide recognition, mRNA and ribosome binding, glycosylation,disulfide bond formation, sugar processing, oligomerization, forexample.

The properties of the insect signal peptide are expected to be moreoptimal for the efficiency of the translation process in insect cellsthan those from vertebrate proteins. This phenomenon can generally beexplained by the fact that proteins secreted from cells are synthesizedas precursor molecules containing hydrophobic N-terminal signalpeptides. The signal peptides direct transport of the select protein toits target membrane and are then cleaved by a peptidase on the membrane,such as the endoplasmic reticulum, when the protein passes through it.

Another exemplary insect signal sequence is the sequence encoding forDrosophila cuticle proteins such as CP1, CP2, CP3 or CP4 (Summers, U.S.Pat. No. 5,278,050; the entirety of which is herein incorporated byreference). Most of a 9 kb region of the Drosophila genome containinggenes for the cuticle proteins has been sequenced. Four of the fivecuticle genes contains a signal peptide coding sequence interrupted by ashort intervening sequence (about 60 base pairs) at a conserved site.Conserved sequences occur in the 5′ mRNA untranslated region, in theadjacent 35 base pairs of upstream flanking sequence and at −200 basepairs from the mRNA start position in each of the cuticle genes.

Standard methods of insect cell culture, cotransfection and preparationof plasmids are set forth in Summers and Smith (Summers and Smith, AManual of Methods for Baculovirus Vectors and Insect Cell CultureProcedures, Texas Agricultural Experiment Station Bulletin No. 1555,Texas A&M University (1987)). Procedures for the cultivation of virusesand cells are described in Volkman and Summers, J. Virol 19:820-832(1975) and Volkman et al., J. Virol 19:820-832 (1976); both of which areherein incorporated by reference in their entirety.

(e) Bacterial Constructs and Transformed Bacterial Cells

The present invention also relates to a bacterial recombinant vectorcomprising exogenous genetic material. The present invention alsorelates to a bacteria cell comprising a bacterial recombinant vector.The present invention also relates to methods for obtaining arecombinant bacteria host cell, comprising introducing into a bacterialhost cell exogenous genetic material.

The bacterial recombinant vector may be any vector which can beconveniently subjected to recombinant DNA procedures. The choice of avector will typically depend on the compatibility of the vector with thebacterial host cell into which the vector is to be introduced. Thevector may be a linear or a closed circular plasmid. The vector systemmay be a single vector or plasmid or two or more vectors or plasmidswhich together contain the total DNA to be introduced into the genome ofthe bacterial host. In addition, the bacterial vector may be anexpression vector. Nucleic acid molecules encoding protein homologues orfragments thereof can, for example, be suitably inserted into areplicable vector for expression in the bacterium under the control of asuitable promoter for bacteria. Many vectors are available for thispurpose and selection of the appropriate vector will depend mainly onthe size of the nucleic acid to be inserted into the vector and theparticular host cell to be transformed with the vector. Each vectorcontains various components depending on its function (amplification ofDNA or expression of DNA) and the particular host cell with which it iscompatible. The vector components for bacterial transformation generallyinclude, but are not limited to, one or more of the following: a signalsequence, an origin of replication, one or more marker genes and aninducible promoter.

In general, plasmid vectors containing replicon and control sequencesthat are derived from species compatible with the host cell are used inconnection with bacterial hosts. The vector ordinarily carries areplication site, as well as marking sequences that are capable ofproviding phenotypic selection in transformed cells. For example, E.coli is typically transformed using pBR322, a plasmid derived from an E.coli species (see, e.g., Bolivar et al., Gene 2:95 (1977); the entiretyof which is herein incorporated by reference). pBR322 contains genes forampicillin and tetracycline resistance and thus provides easy means foridentifying transformed cells. The pBR322 plasmid, or other microbialplasmid or phage, also generally contains, or is modified to contain,promoters that can be used by the microbial organism for expression ofthe selectable marker genes.

Nucleic acid molecules encoding protein or fragments thereof may beexpressed not only directly, but also as a fusion with anotherpolypeptide, preferably a signal sequence or other polypeptide having aspecific cleavage site at the N-terminus of the mature polypeptide. Ingeneral, the signal sequence may be a component of the vector, or it maybe a part of the polypeptide DNA that is inserted into the vector. Theheterologous signal sequence selected should be one that is recognizedand processed (i.e., cleaved by a signal peptidase) by the host cell.For bacterial host cells that do not recognize and process the nativepolypeptide signal sequence, the signal sequence is substituted by abacterial signal sequence selected, for example, from the groupconsisting of the alkaline phosphatase, penicillinase, lpp, orheat-stable enterotoxin II leaders.

Both expression and cloning vectors contain a nucleic acid sequence thatenables the vector to replicate in one or more selected host cells.Generally, in cloning vectors this sequence is one that enables thevector to replicate independently of the host chromosomal DNA andincludes origins of replication or autonomously replicating sequences.Such sequences are well known for a variety of bacteria. The origin ofreplication from the plasmid pBR322 is suitable for most Gram-negativebacteria.

Expression and cloning vectors also generally contain a selection gene,also termed a selectable marker. This gene encodes a protein necessaryfor the survival or growth of transformed host cells grown in aselective culture medium. Host cells not transformed with the vectorcontaining the selection gene will not survive in the culture medium.Typical selection genes encode proteins that (a) confer resistance toantibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate,or tetracycline, (b) complement auxotrophic deficiencies, or (c) supplycritical nutrients not available from complex media, e.g., the geneencoding D-alanine racemase for Bacilli. One example of a selectionscheme utilizes a drug to arrest growth of a host cell. Those cells thatare successfully transformed with a heterologous protein homologue orfragment thereof produce a protein conferring drug resistance and thussurvive the selection regimen.

The expression vector for producing a protein or fragment thereof canalso contains an inducible promoter that is recognized by the hostbacterial organism and is operably linked to the nucleic acid encoding,for example, the nucleic acid molecule encoding the protein homologue orfragment thereof of interest. Inducible promoters suitable for use withbacterial hosts include the β-lactamase and lactose promoter systems(Chang et al., Nature 275:615 (1978); Goeddel et al., Nature 281:544(1979); both of which are herein incorporated by reference in theirentirety), the arabinose promoter system (Guzman et al., J. Bacteriol.174:7716-7728 (1992); the entirety of which is herein incorporated byreference), alkaline phosphatase, a tryptophan (trp) promoter system(Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both of which areherein incorporated by reference in their entirety) and hybrid promoterssuch as the tac promoter (deBoer et al., Proc. Natl. Acad. Sci. (USA)80:21-25 (1983); the entirety of which is herein incorporated byreference). However, other known bacterial inducible promoters aresuitable (Siebenlist et al., Cell 20:269 (1980); the entirety of whichis herein incorporated by reference).

Promoters for use in bacterial systems also generally contain aShine-Dalgarno (S.D.) sequence operably linked to the DNA encoding thepolypeptide of interest. The promoter can be removed from the bacterialsource DNA by restriction enzyme digestion and inserted into the vectorcontaining the desired DNA.

Construction of suitable vectors containing one or more of theabove-listed components employs standard ligation techniques. Isolatedplasmids or DNA fragments are cleaved, tailored and re-ligated in theform desired to generate the plasmids required. Examples of availablebacterial expression vectors include, but are not limited to, themultifunctional E. coli cloning and expression vectors such asBluescript™ (Stratagene, La Jolla, Calif.), in which, for example,encoding an A. nidulans protein homologue or fragment thereof homologue,may be ligated into the vector in frame with sequences for theamino-terminal Met and the subsequent 7 residues of β-galactosidase sothat a hybrid protein is produced; pIN vectors (Van Heeke and Schuster,J. Biol. Chem. 264:5503-5509 (1989), the entirety of which is hereinincorporated by reference); and the like. pGEX vectors (Promega, MadisonWis. U.S.A.) may also be used to express foreign polypeptides as fusionproteins with glutathione S-transferase (GST). In general, such fusionproteins are soluble and can easily be purified from lysed cells byadsorption to glutathione-agarose beads followed by elution in thepresence of free glutathione. Proteins made in such systems are designedto include heparin, thrombin or factor XA protease cleavage sites sothat the cloned polypeptide of interest can be released from the GSTmoiety at will.

Suitable host bacteria for a bacterial vector include archaebacteria andeubacteria, especially eubacteria and most preferablyEnterobacteriaceae. Examples of useful bacteria include Escherichia,Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsiella,Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla andParacoccus. Suitable E. coli hosts include E. coli W3110 (American TypeCulture Collection (ATCC) 27,325, Manassas, Va. U.S.A.), E. coli 294(ATCC 31,446), E. coli B and E. coli X1776 (ATCC 31,537). These examplesare illustrative rather than limiting. Mutant cells of any of theabove-mentioned bacteria may also be employed. It is, of course,necessary to select the appropriate bacteria taking into considerationreplicability of the replicon in the cells of a bacterium. For example,E. coli, Serratia, or Salmonella species can be suitably used as thehost when well known plasmids such as pBR322, pBR325, pACYC177, orpKN410 are used to supply the replicon. E. coli strain W3110 is apreferred host or parent host because it is a common host strain forrecombinant DNA product fermentations. Preferably, the host cell shouldsecrete minimal amounts of proteolytic enzymes.

Host cells are transfected and preferably transformed with theabove-described vectors and cultured in conventional nutrient mediamodified as appropriate for inducing promoters, selecting transformants,or amplifying the genes encoding the desired sequences.

Numerous methods of transfection are known to the ordinarily skilledartisan, for example, calcium phosphate and electroporation. Dependingon the host cell used, transformation is done using standard techniquesappropriate to such cells. The calcium treatment employing calciumchloride, as described in section 1.82 of Sambrook et al., MolecularCloning: A Laboratory Manual, New York: Cold Spring Harbor LaboratoryPress, (1989), is generally used for bacterial cells that containsubstantial cell-wall barriers. Another method for transformationemploys polyethylene glycol/DMSO, as described in Chung and Miller(Chung and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety ofwhich is herein incorporated by reference). Yet another method is theuse of the technique termed electroporation.

Bacterial cells used to produce the polypeptide of interest for purposesof this invention are cultured in suitable media in which the promotersfor the nucleic acid encoding the heterologous polypeptide can beartificially induced as described generally, e.g., in Sambrook et al.,Molecular Cloning: A Laboratory Manual, New York: Cold Spring HarborLaboratory Press, (1989). Examples of suitable media are given in U.S.Pat. Nos. 5,304,472 and 5,342,763; both of which are incorporated byreference in their entirety.

In addition to the above discussed procedures, practitioners arefamiliar with the standard resource materials which describe specificconditions and procedures for the construction, manipulation andisolation of macromolecules (e.g., DNA molecules, plasmids, etc.),generation of recombinant organisms and the screening and isolating ofclones, (see for example, Sambrook et al., Molecular Cloning: ALaboratory Manual, Cold Spring Harbor Press (1989); Mailga et al.,Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995), theentirety of which is herein incorporated by reference; Birren et al.,Genome Analysis: Analyzing DNA, 1, Cold Spring Harbor, N.Y., theentirety of which is herein incorporated by reference).

(f) Computer Readable Media

The nucleotide sequence provided in SEQ ID NO: 1 through SEQ ID NO: 3204or fragment thereof, or complement thereof, or a nucleotide sequence atleast 90% identical, preferably 95%, identical even more preferably 99%or 100% identical to the sequence provided in SEQ ID NO: 1 through SEQID NO: 3204 or fragment thereof, or complement thereof, can be“provided” in a variety of mediums to facilitate use. Such a medium canalso provide a subset thereof in a form that allows a skilled artisan toexamine the sequences.

A preferred subset of nucleotide sequences are those nucleic acidsequences that encode a maize or soybean methionine adenosyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or soybean S-adenosylmethioninedecarboxylase enzyme or complement thereof or fragment of either, anucleic acid molecule that encodes a maize or soybean aspartate kinaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or soybean aspartate-semialdehydedehydrogenase enzyme or complement thereof or fragment of either, anucleic acid molecule that encodes a maize or soybeanO-succinylhomoserine (thiol)-lyase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize orsoybean cystathionine β-lyase enzyme or complement thereof or fragmentof either, a nucleic acid molecule that encodes a maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-5-methyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or a soybean adenosylhomocysteinase enzymeor complement thereof or fragment of either, a nucleic acid moleculethat encodes a maize or a soybean cystathionine β-synthase enzyme orcomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean cytsathionine γ-lyase enzyme or complementthereof or fragment of either and a nucleic acid molecule that encodes amaize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complementthereof or fragment of either.

A further preferred subset of nucleic acid sequences is where the subsetof sequences is two proteins or fragments thereof, more preferably threeproteins or fragments thereof and even more preferable four proteins orfragments thereof, these nucleic acid sequences are selected from thegroup that comprises a maize or soybean methionine adenosyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or soybean S-adenosylmethioninedecarboxylase enzyme or complement thereof or fragment of either, anucleic acid molecule that encodes a maize or soybean aspartate kinaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or soybean aspartate-semialdehydedehydrogenase enzyme or complement thereof or fragment of either, anucleic acid molecule that encodes a maize or soybeanO-succinylhomoserine (thiol)-lyase enzyme or complement thereof orfragment of either, a nucleic acid molecule that encodes a maize orsoybean cystathionine β-lyase enzyme or complement thereof or fragmentof either, a nucleic acid molecule that encodes a maize or soybean5-methyltetrahydropteroyltriglutamate-homocysteine-S-methyltransferaseenzyme or complement thereof or fragment of either, a nucleic acidmolecule that encodes a maize or a soybean adenosylhomocysteinase enzymeor complement thereof or fragment of either, a nucleic acid moleculethat encodes a maize or a soybean cystathionine β-synthase enzyme orcomplement thereof or fragment of either, a nucleic acid molecule thatencodes a maize or a soybean cytsathionine γ-lyase enzyme or complementthereof or fragment of either and a nucleic acid molecule that encodes amaize or a soybean O-acetylhomoserine (thiol)-lyase enzyme or complementthereof or fragment of either.

In one application of this embodiment, a nucleotide sequence of thepresent invention can be recorded on computer readable media. As usedherein, “computer readable media” refers to any medium that can be readand accessed directly by a computer. Such media include, but are notlimited to: magnetic storage media, such as floppy discs, hard disc,storage medium and magnetic tape: optical storage media such as CD-ROM;electrical storage media such as RAM and ROM; and hybrids of thesecategories such as magnetic/optical storage media. A skilled artisan canreadily appreciate how any of the presently known computer readablemediums can be used to create a manufacture comprising computer readablemedium having recorded thereon a nucleotide sequence of the presentinvention.

As used herein, “recorded” refers to a process for storing informationon computer readable medium. A skilled artisan can readily adopt any ofthe presently known methods for recording information on computerreadable medium to generate media comprising the nucleotide sequenceinformation of the present invention. A variety of data storagestructures are available to a skilled artisan for creating a computerreadable medium having recorded thereon a nucleotide sequence of thepresent invention. The choice of the data storage structure willgenerally be based on the means chosen to access the stored information.In addition, a variety of data processor programs and formats can beused to store the nucleotide sequence information of the presentinvention on computer readable medium. The sequence information can berepresented in a word processing text file, formatted incommercially-available software such as WordPerfect and Microsoft Word,or represented in the form of an ASCII file, stored in a databaseapplication, such as DB2, Sybase, Oracle, or the like. A skilled artisancan readily adapt any number of data processor structuring formats (e.g.text file or database) in order to obtain computer readable mediumhaving recorded thereon the nucleotide sequence information of thepresent invention.

By providing one or more of nucleotide sequences of the presentinvention, a skilled artisan can routinely access the sequenceinformation for a variety of purposes. Computer software is publiclyavailable which allows a skilled artisan to access sequence informationprovided in a computer readable medium. The examples which followdemonstrate how software which implements the BLAST (Altschul et al., J.Mol. Biol. 215:403-410 (1990), the entirety of which is hereinincorporated by reference) and BLAZE (Brutlag et al., Comp. Chem.17:203-207 (1993), the entirety of which is herein incorporated byreference) search algorithms on a Sybase system can be used to identifyopen reading frames (ORFs) within the genome that contain homology toORFs or proteins from other organisms. Such ORFs are protein-encodingfragments within the sequences of the present invention and are usefulin producing commercially important proteins such as enzymes used inamino acid biosynthesis, metabolism, transcription, translation, RNAprocessing, nucleic acid and a protein degradation, protein modificationand DNA replication, restriction, modification, recombination andrepair.

The present invention further provides systems, particularlycomputer-based systems, which contain the sequence information describedherein. Such systems are designed to identify commercially importantfragments of the nucleic acid molecule of the present invention. As usedherein, “a computer-based system” refers to the hardware means, softwaremeans and data storage means used to analyze the nucleotide sequenceinformation of the present invention. The minimum hardware means of thecomputer-based systems of the present invention comprises a centralprocessing unit (CPU), input means, output means and data storage means.A skilled artisan can readily appreciate that any one of the currentlyavailable computer-based system are suitable for use in the presentinvention.

As indicated above, the computer-based systems of the present inventioncomprise a data storage means having stored therein a nucleotidesequence of the present invention and the necessary hardware means andsoftware means for supporting and implementing a search means. As usedherein, “data storage means” refers to memory that can store nucleotidesequence information of the present invention, or a memory access meanswhich can access manufactures having recorded thereon the nucleotidesequence information of the present invention. As used herein, “searchmeans” refers to one or more programs which are implemented on thecomputer-based system to compare a target sequence or target structuralmotif with the sequence information stored within the data storagemeans. Search means are used to identify fragments or regions of thesequence of the present invention that match a particular targetsequence or target motif. A variety of known algorithms are disclosedpublicly and a variety of commercially available software for conductingsearch means are available can be used in the computer-based systems ofthe present invention. Examples of such software include, but are notlimited to, MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of theavailable algorithms or implementing software packages for conductinghomology searches can be adapted for use in the present computer-basedsystems.

The most preferred sequence length of a target sequence is from about 10to 100 amino acids or from about 30 to 300 nucleotide residues. However,it is well recognized that during searches for commercially importantfragments of the nucleic acid molecules of the present invention, suchas sequence fragments involved in gene expression and proteinprocessing, may be of shorter length.

As used herein, “a target structural motif,” or “target motif,” refersto any rationally selected sequence or combination of sequences in whichthe sequences the sequence(s) are chosen based on a three-dimensionalconfiguration which is formed upon the folding of the target motif.There are a variety of target motifs known in the art. Protein targetmotifs include, but are not limited to, enzymatic active sites andsignal sequences. Nucleic acid target motifs include, but are notlimited to, promoter sequences, cis elements, hairpin structures andinducible expression elements (protein binding sequences).

Thus, the present invention further provides an input means forreceiving a target sequence, a data storage means for storing the targetsequences of the present invention sequence identified using a searchmeans as described above and an output means for outputting theidentified homologous sequences. A variety of structural formats for theinput and output means can be used to input and output information inthe computer-based systems of the present invention. A preferred formatfor an output means ranks fragments of the sequence of the presentinvention by varying degrees of homology to the target sequence ortarget motif. Such presentation provides a skilled artisan with aranking of sequences which contain various amounts of the targetsequence or target motif and identifies the degree of homology containedin the identified fragment.

A variety of comparing means can be used to compare a target sequence ortarget motif with the data storage means to identify sequence fragmentssequence of the present invention. For example, implementing softwarewhich implement the BLAST and BLAZE algorithms (Altschul et al., J. Mol.Biol. 215:403-410 (1990)) can be used to identify open frames within thenucleic acid molecules of the present invention. A skilled artisan canreadily recognize that any one of the publicly available homology searchprograms can be used as the search means for the computer-based systemsof the present invention.

Having now generally described the invention, the same will be morereadily understood through reference to the following examples which areprovided by way of illustration and are not intended to be limiting ofthe present invention, unless specified.

EXAMPLE 1

The MONN01 cDNA library is a normalized library generated from maize(DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) total leaf tissue at theV6 plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedwhen the maize plant is at the 6-leaf development stage. The older, morejuvenile leaves, which are in a basal position, as well as the younger,more adult leaves, which are more apical are cut at the base of theleaves. The leaves are then pooled and immediately transferred to liquidnitrogen containers in which the pooled leaves are crushed. Theharvested tissue is then stored at −80° C. until RNA preparation.

The SATMON001 cDNA library is generated from maize (B73, IllinoisFoundation Seeds, Champaign, Ill. U.S.A.) immature tassels at the V6plant development stage. Seeds are planted at a depth of approximately 3cm into 2-3 inch peat pots containing Metro 200 growing medium. After2-3 weeks growth they are transplanted into 10 inch pots containing thesame growing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Tissue from the maize plant is collected atthe V6 stage. At that stage the tassel is an immature tassel of about2-3 cm in length. The tassels are removed and frozen in liquid nitrogen.The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON003 library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign, Ill. U.S.A.) roots at the V6 developmentalstage. Seeds are planted at a depth of approximately 3 cm in coil into2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeksgrowth, the seedlings are transplanted into 10 inch pots containing theMetro 200 growing medium. Plants are watered daily beforetransplantation and approximately 3 times a week after transplantation.Peters 15-16-17 fertilizer is applied approximately three times per weekafter transplanting at a concentration of 150 ppm N. Two to three timesduring the life time of the plant from transplanting to flowering atotal of approximately 900 mg Fe is added to each pot. Maize plants aregrown in the green house in approximately 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Tissue is collected when the maize plant isat the 6 leaf development stage. The root system is cut from maize plantand washed with water to free it from the soil. The tissue is thenimmediately frozen in liquid nitrogen. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SATMON004 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign, Ill. U.S.A.) total leaf tissue at the V6plant development stage. Seeds are planted at a depth of approximately 3cm into 2-3 inch peat pots containing Metro 200 growing medium. After2-3 weeks growth they are transplanted into 10 inch pots containing thesame growing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Tissue is collected when the maize plant isat the 6-leaf development stage. The older, more juvenile leaves, whichare in a basal position, as well as the younger, more adult leaves,which are more apical are cut at the base of the leaves. The leaves arethen pooled and immediately transferred to liquid nitrogen containers inwhich the pooled leaves are crushed. The harvested tissue is then storedat −80° C. until RNA preparation.

The SATMON005 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign Ill., U.S.A.) root tissue at the V6development stage. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. Tissue is collected when themaize plant is at the 6-leaf development stage. The root system is cutfrom the mature maize plant and washed with water to free it from thesoil. The tissue is immediately frozen in liquid nitrogen and theharvested tissue is then stored at −80° C. until RNA preparation.

The SATMON006 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign Ill., U.S.A.) total leaf tissue at the V6plant development stage. Seeds are planted at a depth of approximately 3cm into 2-3 inch peat pots containing Metro 200 growing medium. After2-3 weeks growth they are transplanted into 10 inch pots containing thesame growing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Tissue is collected when the maize plant isat the 6-leaf development stage. The older more juvenile leaves, whichare in a basal position, as well as the younger more adult leaves, whichare more apical are cut at the base of the leaves. The leaves are thenpooled and immediately transferred to liquid nitrogen containers inwhich the pooled leaves are crushed. The harvested tissue is then storedat −80° C. until RNA preparation.

The SATMON007 cDNA library is generated from the primary root tissue of5 day old maize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings.Seeds are planted on a moist filter paper on a covered tray that is keptin the dark until germination (one day). After germination, the trays,along with the moist paper, are moved to a greenhouse where the maizeplants are grown in the greenhouse in 15 hr day/9 hr night cycles forapproximately 5 days. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. The primary roottissue is collected when the seedlings are 5 days old. At this stage,the primary root (radicle) is pushed through the coleorhiza which itselfis pushed through the seed coat. The primary root, which is about 2-3 cmlong, is cut and immediately frozen in liquid nitrogen and then storedat −80° C. until RNA preparation.

The SATMON008 cDNA library is generated from the primary shoot(coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb, Ill.U.S.A.) seedlings which are approximately 5 days old. Seeds are plantedon a moist filter paper on a covered tray that is kept in the dark untilgermination (one day). Then the trays containing the seeds are moved toa greenhouse at 15 hr daytime/9 hr nighttime cycles and grown until theyare 5 days post germination. The daytime temperature is approximately80° F. and the nighttime temperature is approximately 70° F. Tissue iscollected when the seedlings are 5 days old. At this stage, the primaryshoot (coleoptile) is pushed through the seed coat and is about 2-3 cmlong. The coleoptile is dissected away from the rest of the seedling,immediately frozen in liquid nitrogen and then stored at −80° C. untilRNA preparation.

The SATMON009 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) leaves at the 8 leaf stage (V8 plantdevelopment stage). Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is 80° F. and the nighttime temperatureis 70° F. Supplemental lighting is provided by 1000 W sodium vaporlamps. Tissue is collected when the maize plant is at the 8-leafdevelopment stage. The older more juvenile leaves, which are in a basalposition, as well as the younger more adult leaves, which are moreapical, are cut at the base of the leaves. The leaves are then pooledand then immediately transferred to liquid nitrogen containers in whichthe pooled leaves are crushed. The harvested tissue is then stored at−80° C. until RNA preparation.

The SATMON010 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) root tissue at the V8 plant developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is 80° F. and the nighttime temperature is 70° F.Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissueis collected when the maize plant is at the V8 development stage. Theroot system is cut from this mature maize plant and washed with water tofree it from the soil. The tissue is immediately frozen in liquidnitrogen. The harvested tissue is then stored at −80° C. until RNApreparation.

The SATMON011 cDNA library is generated from undeveloped maize (DK604,Dekalb Genetics, Dekalb, Ill. U.S.A.) leaf at the V6 plant developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is approximately 80° F. and the nighttime temperature isapproximately 70° F. Supplemental lighting is provided by 1000 W sodiumvapor lamps. Tissue is collected when the maize plant is at the 6-leafdevelopment stage. The second youngest leaf which is at the base of theapical leaf of V6 stage maize plant is cut at the base and immediatelytransferred to liquid nitrogen containers in which the leaf is crushed.The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON012 cDNA library is generated from 2 day post germinationmaize (DK604, Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds areplanted on a moist filter paper on a covered tray that is kept in thedark until germination (one day). Then the trays containing the seedsare moved to the greenhouse and grown at 15 hr daytime/9 hr nighttimecycles until 2 days post germination. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Tissue is collected when the seedlings are 2 days old. At the two daystage, the coleorhiza is pushed through the seed coat and the primaryroot (the radicle) is pierced the coleorhiza but is barely visible.Also, at this two day stage, the coleoptile is just emerging from theseed coat. The 2 days post germination seedlings are then immersed inliquid nitrogen and crushed. The harvested tissue is stored at −80° C.until preparation of total RNA.

The SATMON013 cDNA library is generated from apical maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) meristem founder at the V4 plantdevelopment stage. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Prior to tissue collection, the plant is atthe 4 leaf stage. The lead at the apex of the V4 stage maize plant isreferred to as the meristem founder. This apical meristem founder iscut, immediately frozen in liquid nitrogen and crushed. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SATMON014 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) endosperm fourteen days afterpollination. Seeds are planted at a depth of approximately 3 cm into 2-3inch peat pots containing Metro 200 growing medium. After 2-3 weeksgrowth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. After the V10 stage, the maize plant earshoots are ready for fertilization. At this stage, the ear shoots areenclosed in a paper bag before silk emergence to withhold the pollen.The ear shoots are pollinated and 14 days after pollination, the earsare pulled out and then the kernels are plucked out of the ears. Eachkernel is then dissected into the embryo and the endosperm and thealeurone layer is removed. After dissection, the endosperms areimmediately frozen in liquid nitrogen and then stored at −80° C. untilRNA preparation.

The SATMON016 library is a maize (DK604, Dekalb Genetics, Dekalb, Ill.U.S.A.) sheath library collected at the V8 developmental stage. Seedsare planted in a depth of approximately 3 cm in solid into 2-3 inch potscontaining Metro growing medium. After 2-3 weeks growth, they aretransplanted into 10″ pots containing the same. Plants are watered dailybefore transplantation and approximately the times a week aftertransplantation. Peters 15-16-17 fertilizer is applied approximatelythree times per week after transplanting, at a strength of 150 ppm N.Two to three times during the life time of the plant from transplantingto flowering, a total of approximately 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 w sodium vapor lamps. When the maize plants are at theV8 stage the 5^(th) and 6^(th) leaves from the bottom exhibit fullydeveloped leaf blades. At the base of these leaves, the ligule isdifferentiated and the leaf blade is joined to the sheath. The sheath isdissected away from the base of the leaf then the sheath is frozen inliquid nitrogen and crushed. The tissue is then stored at −80° C. untilRNA preparation.

The SATMON017 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) embryo seventeen days after pollination.Seeds are planted at a depth of approximately 3 cm into 2-3 inch peatpots containing Metro 200 growing medium. After 2-3 weeks growth theseeds are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is approximately 80° F. and the nighttime temperature isapproximately 70° F. Supplemental lighting is provided by 1000 W sodiumvapor lamps. After the V10 stage, the ear shoots of maize plant, whichare ready for fertilization, are enclosed in a paper bag before silkemergence to withhold the pollen. The ear shoots are fertilized and 21days after pollination, the ears are pulled out and the kernels areplucked out of the ears. Each kernel is then dissected into the embryoand the endosperm and the aleurone layer is removed. After dissection,the embryos are immediately frozen in liquid nitrogen and then stored at−80° C. until RNA preparation.

The SATMON019 (Lib3054) cDNA library is generated from maize (DK604,Dekalb Genetics, Dekalb, Ill. U.S.A.) culm (stem) at the V8developmental stage. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. When the maize plant is at the V8stage, the 5th and 6th leaves from the bottom have fully developed leafblades. The region between the nodes of the 5th and the sixth leavesfrom the bottom is the region of the stem that is collected. The leavesare pulled out and the sheath is also torn away from the stem. This stemtissue is completely free of any leaf and sheath tissue. The stem tissueis then frozen in liquid nitrogen and stored at −80° C. until RNApreparation.

The SATMON020 cDNA library is from a maize (DK604, Dekalb Genetics,Dekalb, Ill. U.S.A.) Hill Type II-Initiated Callus. Petri platescontaining approximately 25 ml of Type II initiation media are prepared.This medium contains N6 salts and vitamins, 3% sucrose, 2.3 g/literproline 0.1 g/liter enzymatic casein hydrolysate, 2 mg/liter2,4-dichloro phenoxy-acetic acid (2,4, D), 15.3 mg/liter AgNO₃ and 0.8%bacto agar and is adjusted to pH 6.0 before autoclaving. At 9-11 daysafter pollination, an ear with immature embryos measuring approximately1-2 mm in length is chosen. The husks and silks are removed and then theear is broken into halves and placed in an autoclaved solution ofClorox/TWEEN 20 sterilizing solution. Then the ear is rinsed withdeionized water. Then each embryo is extracted from the kernel. Intactembryos are placed in contact with the medium, scutellar side up).Multiple embryos are plated on each plate and the plates are incubatedin the dark at 25° C. Type II calluses are friable, can be subculturedwith a spatula, frequently regenerate via somatic embryogenesis and arerelatively undifferentiated. As seen in the microscope, the Tape IIcalluses show color ranging from translucent to light yellow andheterogeneity on with respect to embryoid structure as well as stage ofembryoid development. Once Type II callus are formed, the calluses istransferred to type II callus maintenance medium without AgNO₃. Every7-10 days, the callus is subcultured. About 4 weeks after embryoisolation the callus is removed from the plates and then frozen inliquid nitrogen. The harvested tissue is stored at −80° C. until RNApreparation.

The SATMON021 cDNA library is generated from the immature maize (DK604,Dekalb Genetics, Dekalb Ill., U.S.A.) tassel at the V8 plant developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is approximately 80° F. and the nighttime temperature isapproximately 70° F. Supplemental lighting is provided by 1000 W sodiumvapor lamps. As the maize plant enters the V8 stage, tassels which are15-20 cm in length are collected and frozen in liquid nitrogen. Theharvested tissue is stored at −80° C. until RNA preparation.

The SATMON022 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) ear (growing silks) at the V8 plantdevelopment stage. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Zea mays plants are grown in the greenhouse in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. Tissue is collected when theplant is in the V8 stage. At this stage, some immature ear shoots arevisible. The immature ear shoots (approximately 1 cm in length) arepulled out, frozen in liquid nitrogen and then stored at −80° C. untilRNA preparation.

The SATMON23 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) ear (growing silk) at the V8 developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the greenhouse in 15 hr day/9 hr night cycles. The daytimetemperature is approximately 80° F. and the nighttime temperature isapproximately 70° F. When the tissue is harvested at the V8 stage, thelength of the ear that is harvested is about 110-15 cm and the silks arejust exposed (approximately 1 inch). The ear along with the silks isfrozen in liquid nitrogen and then the tissue is stored at −80° C. untilRNA preparation.

The SATMON024 cDNA library is generated from the immature maize (DK604,Dekalb Genetics, Dekalb, Ill. U.S.A.) tassel at the V9 developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is approximately 80° F. and the nighttime temperature isapproximately 70° F. As a maize plant enters the V9 stage, the tassel israpidly developing and a 37 cm tassel along with the glume, anthers andpollen is collected and frozen in liquid nitrogen. The harvested tissueis stored at −80° C. until RNA preparation.

The SATMON025 cDNA library is from maize (DK604, Dekalb Genetics,Dekalb, Ill. U.S.A.) Hill Type II-Regenerated Callus. Type II callus isgrown in initiation media as described for SATMON020 and then theembryoids on the surface of the Type II callus are allowed to mature andgerminate. The 1-2 gm fresh weight of the soft friable type calluscontaining numerous embryoids are transferred to 100×15 mm petri platescontaining 25 ml of regeneration media. Regeneration media consists ofMurashige and Skoog (MS) basal salts, modified White's vitamins (0.2g/liter glycine and 0.5 g/liter myo-inositol and 0.8% bacto agar(6SMS0D)). The plates are then placed in the dark after covering withparafilm. After 1 week, the plates are moved to a lighted growth chamberwith 16 hr light and 8 hr dark photoperiod. Three weeks after platingthe Type II callus to 6SMS0D, the callus exhibit shoot formation. Thecallus and the shoots are transferred to fresh 6SMS0D plates for another2 weeks. The callus and the shoots are then transferred to petri plateswith reduced sucrose (3SMS0D). Upon distinct formation of a root andshoot, the newly developed green plants are then removed out with aspatula and frozen in liquid nitrogen containers. The harvested tissueis then stored at −80° C. until RNA preparation.

The SATMON026 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) juvenile/adult shift leaves at the V8plant development stage. Seeds are planted at a depth of approximately 3cm into 2-3 inch peat pots containing Metro 200 growing medium. After2-3 weeks growth they are transplanted into 10 inch pots containing thesame growing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. Tissue is collected when themaize plants are at the 8-leaf development stage. Leaves are foundedsequentially around the meristem over weeks of time and the older, morejuvenile leaves arise earlier and in a more basal position than theyounger, more adult leaves, which are in a more apical position. In a V8plant, some leaves which are in the middle portion of the plant exhibitcharacteristics of both juvenile as well as adult leaves. They exhibit ayellowing color but also exhibit, in part, a green color. These leavesare termed juvenile/adult shift leaves. The juvenile/adult shift leaves(the 4th, 5th leaves from the bottom) are cut at the base, pooled andtransferred to liquid nitrogen in which they are then crushed. Theharvested tissue is then stored at −80° C. until RNA preparation.

The SATMON027 cDNA library is generated from 6 day maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) leaves. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the Metro 200 growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Zea mays plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Prior to tissuecollection, when the plant is at the 8-leaf stage, water is held backfor six days. The older, more juvenile leaves, which are in a basalposition, as well as the younger, more adult leaves, which are moreapical, are all cut at the base of the leaves. All the leaves exhibitsignificant wilting. The leaves are then pooled and immediatelytransferred to liquid nitrogen containers in which the pooled leaves arethen crushed. The harvested tissue is then stored at −80° C. until RNApreparation.

The SATMON028 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) roots at the V8 developmental stage thatare subject to six days water stress. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the Metro 200 growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Prior to tissuecollection, when the plant is at the 8-leaf stage, water is held backfor six days. The root system is cut, shaken and washed to remove soil.Root tissue is then pooled and immediately transferred to liquidnitrogen containers in which the pooled leaves are then crushed. Theharvested tissue is then stored at −80° C. until RNA preparation.

The SATMON029 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) seedlings at the etiolated stage. Seedsare planted on a moist filter paper on a covered tray that is kept inthe dark for 4 days at approximately 70° F. Tissue is collected when theseedlings are 4 days old. By 4 days, the primary root has penetrated thecoleorhiza and is about 4-5 cm and the secondary lateral roots have alsomade their appearance. The coleoptile has also pushed through the seedcoat and is about 4-5 cm long. The seedlings are frozen in liquidnitrogen and crushed. The harvested tissue is then stored at −80° C.until RNA preparation.

The SATMON030 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) root tissue at the V4 plant developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growth,they are transplanted into 10 inch pots containing the same. Plants arewatered daily before transplantation and approximately 3 times a weekafter transplantation. Peters 15-16-17 fertilizer is appliedapproximately three times per week after transplanting, at a strength of150 ppm N. Two to three times during the life time of the plant, fromtransplanting to flowering, a total of approximately 900 mg Fe is addedto each pot. Maize plants are grown in the green house in 15 hr day/9 hrnight cycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 sodium vapor lamps. Tissue is collected when the maizeplant is at the 4 leaf development stage. The root system is cut fromthe mature maize plant and washed with water to free it from the soil.The tissue is then immediately frozen in liquid nitrogen. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SATMON031 cDNA library is generated from the maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) leaf tissue at the V4 plant developmentstage. Seeds are planted at a depth of approximately 3 cm into 2-3 inchpeat pots containing Metro 200 growing medium. After 2-3 weeks growththey are transplanted into 10 inch pots containing the same growingmedium. Plants are watered daily before transplantation and three timesa week after transplantation. Peters 15-16-17 fertilizer is appliedthree times per week after transplanting at a strength of 150 ppm N. Twoto three times during the lifetime of the plant, from transplanting toflowering, a total of 900 mg Fe is added to each pot. Maize plants aregrown in the green house in 15 hr day/9 hr night cycles. The daytimetemperature is 80° F. and the nighttime temperature is 70° F.Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissueis collected when the maize plant is at the 4-leaf development stage.The third leaf from the bottom is cut at the base and immediately frozenin liquid nitrogen and crushed. The tissue is immediately frozen inliquid nitrogen. The harvested tissue is then stored at −80° C. untilRNA preparation.

The SATMON033 cDNA library is generated from maize (DK604, DekalbGenetics, Dekalb, Ill. U.S.A.) embryo tissue 13 days after pollination.Seeds are planted at a depth of approximately 3 cm into 2-3 inch peatpots containing Metro 200 growing medium. After 2-3 weeks growth theyare transplanted into 10 inch pots containing the same growing medium.Plants are watered daily before transplantation and three times a weekafter transplantation. Peters 15-16-17 fertilizer is applied three timesper week after transplanting at a strength of 150 ppm N. Two to threetimes during the lifetime of the plant, from transplanting to flowering,a total of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Afterthe V10 stage, the ear shoots of the maize plant, which are ready forfertilization, are enclosed in a paper bag before silk emergent towithhold the pollen. The ear shoots are pollinated and 13 days afterpollination, the ears are pulled out and then the kernels are pluckedcut of the ears. Each kernel is then dissected into the embryo and theendosperm and the aleurone layer is removed. After dissection, theembryos are immediately frozen in liquid nitrogen and then stored at−80° C. until RNA preparation.

The SATMON034 cDNA library is generated from cold stressed maize (DK604,Dekalb Genetics, Dekalb, Ill. U.S.A.) seedlings. Seeds are planted on amoist filter paper on a covered tray that is kept on at 110° C. for 7days. After 7 days, the temperature is shifted to 15° C. for one dayuntil germination of the seed. Tissue is collected once the seedlingsare 1 day old. At this point, the coleorhiza has just pushed out of theseed coat and the primary root is just making its appearance. Thecoleoptile has not yet pushed completely through the seed coat and isalso just making its appearance. These 1 day old cold stressed seedlingsare frozen in liquid nitrogen and crushed. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SATMON˜001 (Lib36, Lib83, Lib84) cDNA library is generated frommaize leaves at the V8 plant development stage. Seeds are planted at adepth of approximately 3 cm into 2-3 inch peat pots containing Metro 200growing medium. After 2-3 weeks growth they are transplanted into 10inch pots containing the same growing medium. Plants are watered dailybefore transplantation and three times a week after transplantation.Peters 15-16-17 fertilizer is applied three times per week aftertransplanting at a strength of 150 ppm N. Two to three times during thelifetime of the plant, from transplanting to flowering, a total of 900mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15hr day/9 hr night cycles. The daytime temperature is approximately 80°F. and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue from the maizeplant is collected at the V8 stage. The older more juvenile leaves in abasal position was well as the younger more adult leaves which are moreapical are all cut at the base, pooled and frozen in liquid nitrogen.The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMONN01 cDNA library is generated from maize (B73, IllinoisFoundation Seeds, Champaign, Ill. U.S.A.) normalized immature tassels atthe V6 plant development stage normalized tissue. Seeds are planted at adepth of approximately 3 cm into 2-3 inch peat pots containing Metro 200growing medium. After 2-3 weeks growth they are transplanted into 10inch pots containing the same growing medium. Plants are watered dailybefore transplantation and three times a week after transplantation.Peters 15-16-17 fertilizer is applied three times per week aftertransplanting at a strength of 150 ppm N. Two to three times during thelifetime of the plant, from transplanting to flowering, a total of 900mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15hr day/9 hr night cycles. The daytime temperature is approximately 80°F. and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue from the maizeplant is collected at the V6 stage. At that stage the tassel is animmature tassel of about 2-3 cm in length. The tassels are removed andfrozen in liquid nitrogen. The harvested tissue is then stored at −80°C. until RNA preparation. Single stranded and double stranded DNArepresenting approximately 1×10⁶ colony forming units are isolated usingstandard protocols. RNA, complementary to the single stranded DNA, issynthesized using the double stranded DNA as a template. BiotinylateddATP is incorporated into the RNA during the synthesis reaction. Thesingle stranded DNA is mixed with the biotinylated RNA in a 1:10 molarratio) and allowed to hybridize. DNA-RNA hybrids are captured onDynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success,N.Y. U.S.A.). The dynabeads with captured hybrids are collected with amagnet. The non-hybridized single stranded molecules remaining afterhybrid capture are converted to double stranded form and represent theprimary normalized library.

The SATMONN04 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign, Ill. U.S.A.) normalized total leaf tissueat the V6 plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedwhen the maize plant is at the 6-leaf development stage. The older, morejuvenile leaves, which are in a basal position, as well as the younger,more adult leaves, which are more apical are cut at the base of theleaves. The leaves are then pooled and immediately transferred to liquidnitrogen containers in which the pooled leaves are crushed. Theharvested tissue is then stored at −80° C. until RNA preparation. Singlestranded and double stranded DNA representing approximately 1×10⁶ colonyforming units are isolated using standard protocols. RNA, complementaryto the single stranded DNA, is synthesized using the double stranded DNAas a template. Biotinylated DATP is incorporated into the RNA during thesynthesis reaction. The single stranded DNA is mixed with thebiotinylated RNA in a 1:10 molar ratio) and allowed to hybridize.DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads withcaptured hybrids are collected with a magnet. The non-hybridized singlestranded molecules remaining after hybrid capture are converted todouble stranded form and represent the primary normalized library.

The SATMONN05 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign Ill., U.S.A.) normalized root tissue at theV6 development stage. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. Tissue is collected when themaize plant is at the 6-leaf development stage. The root system is cutfrom the mature maize plant and washed with water to free it from thesoil. The tissue is immediately frozen in liquid nitrogen and theharvested tissue is then stored at −80° C. until RNA preparation. Thesingle stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio) and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. The non-hybridizedsingle stranded molecules remaining after hybrid capture are convertedto double stranded form and represent the primary normalized library.

The SATMONN06 cDNA library is generated from maize (B73×Mo17, IllinoisFoundation Seeds, Champaign Ill., U.S.A.) normalized total leaf tissueat the V6 plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedwhen the maize plant is at the 6-leaf development stage. The older morejuvenile leaves, which are in a basal position, as well as the youngermore adult leaves, which are more apical are cut at the base of theleaves. The leaves are then pooled and immediately transferred to liquidnitrogen containers in which the pooled leaves are crushed. Theharvested tissue is then stored at −80° C. until RNA preparation. Singlestranded and double stranded DNA representing approximately 1×10⁶ colonyforming units are isolated using standard protocols. RNA, complementaryto the single stranded DNA, is synthesized using the double stranded DNAas a template. Biotinylated dATP is incorporated into the RNA during thesynthesis reaction. The single stranded DNA is mixed with thebiotinylated RNA in a 1:10 molar ratio) and allowed to hybridize.DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads,Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeads withcaptured hybrids are collected with a magnet. The non-hybridized singlestranded molecules remaining after hybrid capture are converted todouble stranded form and represent the primary normalized library.

The CMZ029 (SATMON036) cDNA library is generated from maize (DK604,Dekalb Genetics, Dekalb, Ill. U.S.A.) endosperm 22 days afterpollination. Seeds are planted at a depth of approximately 3 cm into 2-3inch peat pots containing Metro 200 growing medium. After 2-3 weeksgrowth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the green house in 15 hr day/9 hr nightcycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. After the V10 stage, the earshoots of the maize plant, which are ready for fertilization, areenclosed in a paper bag before silk emergent to withhold the pollen. Theear shoots are pollinated and 22 days after pollination, the ears arepulled out and then the kernels are plucked out of the ears. Each kernelis then dissected into the embryo and the endosperm and the aluronelayer is removed. After dissection, the endosperms are immediatelyfrozen in liquid nitrogen and then stored at −80° C. until RNApreparation.

The CMz030 (Lib143) cDNA library is generated from maize seedling tissuetwo days post germination. Seeds are planted on a moist filter paper ona covered try that is keep in the dark until germination. The trays arethen moved to the bench top at 15 hr daytime/9 hr nighttime cycles for 2days post-germination. The day time temperature is 80° F. and thenighttime temperature is 70° F. Tissue is collected when the seedlingsare 2 days old. At this stage, the colehrhiza has pushed through theseed coat and the primary root (the radicle) is just piercing thecolehrhiza and is barely visible. The seedlings are placed at 42° C. for1 hour. Following the heat shock treatment, the seedlings are immersedin liquid nitrogen and crushed. The harvested tissue is stored at −80°until RNA preparation.

The CMz031 (Lib 148) cDNA library is generated from maize pollen tissueat the V10+ plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedfrom V10+ stage plants. The ear shoots, which are ready forfertilization, are enclosed in a paper bag to withhold pollen.Twenty-one days after pollination, prior to removing the ears, the paperbag is shaken to collect the mature pollen. The mature pollen isimmediately frozen in liquid nitrogen containers and the pollen iscrushed. The harvested tissue is then stored at −80° C. until RNApreparation.

The CMz033 (Lib189) cDNA library is generated from maize pooled leaftissue. Samples are harvested from open pollinated plants. Tissue iscollected from maize leaves at the anthesis stage. The leaves arecollect from 10-12 plants and frozen in liquid nitrogen. The harvestedtissue is then stored at −80° C. until RNA preparation.

The CMz034 (Lib3060) cDNA library is generated from maize mature tissueat 40 days post pollination plant development stage. Seeds are plantedat a depth of approximately 3 cm into 2-3 inch peat pots containingMetro 200 growing medium. After 2-3 weeks growth they are transplantedinto 10 inch pots containing the same growing medium. Plants are watereddaily before transplantation and three times a week aftertransplantation. Peters 15-16-17 fertilizer is applied three times perweek after transplanting at a strength of 150 ppm N. Two to three timesduring the lifetime of the plant, from transplanting to flowering, atotal of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected from leaves located two leaves below the ear leaf.This sample represents those genes expressed during onset and earlystages of leaf senescence. The leaves are pooled and immediatelytransferred to liquid nitrogen. The harvested tissue is then stored at−80° C. until RNA preparation.

The CMz036 (Lib3062) cDNA library is generated from maize husk tissue atthe 8 week old plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedfrom 8 week old plants. The husk is separated from the ear andimmediately transferred to liquid nitrogen containers. The harvestedtissue is then stored at −80° C. until RNA preparation.

The CMz037 (Lib3059) cDNA library is generated from maize pooled kernalat 12-15 days after pollienation plant development stage. Sample werecollected from field grown material. Whole kernals from hand pollinated(control pollination) are harvested as whole ears and immediately frozenon dry ice. Kernels from 10-12 ears were pooled and ground together inliquid nitrogen. The harvested tissue is then stored at −80° C. untilRNA preparation.

The CMz039 (Lib3066) cDNA library is generated from maize immatureanther tissue at the 7 week old immature tassel stage. Seeds are plantedat a depth of approximately 3 cm into 2-3 inch peat pots containingMetro 200 growing medium. After 2-3 weeks growth they are transplantedinto 10 inch pots containing the same growing medium. Plants are watereddaily before transplantation and three times a week aftertransplantation. Peters 15-16-17 fertilizer is applied three times perweek after transplanting at a strength of 150 ppm N. Two to three timesduring the lifetime of the plant, from transplanting to flowering, atotal of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected when the maize plant is at the 7 week old immaturetassel stage. At this stage, prior to anthesis, the immature anthers aregreen and enclosed in the staminate spikelet. The developing anthers aredissected away from the 7 week old immature tassel and immediatelyfrozen in liquid nitrogen. The harvested tissue is then stored at −80°C. until RNA preparation.

The CMz040 (Lib3067) cDNA library is generated from maize kernel tissueat the V10+ plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedfrom V10+ stage plants. The ear shoots, which are ready forfertilization, are enclosed in a paper bag before silk emergence towithhold pollen. Five to eight days after controlled pollination. Theears are pulled and the kernels removed. The kernels are immediatelyfrozen in liquid nitrogen. The harvested kernels tissue is then storedat −80° C. until RNA preparation. This sample represents gene expressedin early kernel development, during periods of cell division, amyloplastbiogenesis and early carbon flow across the material to filial tissue.

The CMz041 (Lib3068) cDNA library is generated from maize pollengerminating silk tissue at the V10+ plant development stage. Seeds areplanted at a depth of approximately 3 cm into 2-3 inch peat potscontaining Metro 200 growing medium. After 2-3 weeks growth they aretransplanted into 10 inch pots containing the same growing medium.Plants are watered daily before transplantation and three times a weekafter transplantation. Peters 15-16-17 fertilizer is applied three timesper week after transplanting at a strength of 150 ppm N. Two to threetimes during the lifetime of the plant, from transplanting to flowering,a total of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected from V10+ stage plants when the ear shoots are readyfor fertilization at the silk emergence stage. The emerging silks arepollinated with an excess of pollen under controlled pollinationconditions in the green house. Eighteen hours after pollination thesilks are removed from the ears and immediately frozen in liquidnitrogen containers. This sample represents genes expressed in bothpollen and silk tissue early in pollination. The harvested tissue isthen stored at −80° C. until RNA preparation.

The CMz042 (Lib3069) cDNA library is generated from maize ear tissueexcessively pollinated at the V10+ plant development stage. Seeds areplanted at a depth of approximately 3 cm into 2-3 inch peat potscontaining Metro 200 growing medium. After 2-3 weeks growth they aretransplanted into 10 inch pots containing the same growing medium.Plants are watered daily before transplantation and three times a weekafter transplantation. Peters 15-16-17 fertilizer is applied three timesper week after transplanting at a strength of 150 ppm N. Two to threetimes during the lifetime of the plant, from transplanting to flowering,a total of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected from V10+ stage plants and the ear shoots which areready for fertilization are at the silk emergence stage. The immatureears are pollinated with an excess of pollen under controlledpollination conditions. Eighteen hours post-pollination, the ears areremoved and immediately transferred to liquid nitrogen containers. Theharvested tissue is then stored at −80° C. until RNA preparation.

The CMz044 (Lib3075) cDNA library is generated from maize microsporetissue at the V10+ plant development stage. Seeds are planted at a depthof approximately 3 cm into 2-3 inch peat pots containing Metro 200growing medium. After 2-3 weeks growth they are transplanted into 10inch pots containing the same growing medium. Plants are watered dailybefore transplantation and three times a week after transplantation.Peters 15-16-17 fertilizer is applied three times per week aftertransplanting at a strength of 150 ppm N. Two to three times during thelifetime of the plant, from transplanting to flowering, a total of 900mg Fe is added to each pot. Maize plants are grown in the greenhouse in15 hr day/9 hr night cycles. The daytime temperature is approximately80° F. and the nighttime temperature is approximately 70° F.Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissueis collected from immature anthers from 7 week old tassels. The immatureanthers are first dissected from the 7 week old tassel with a scalpel ona glass slide covered with water. The microspores (immature pollen) arereleased into the water and are recovered by centrifugation. Themicrospore suspension is immediately frozen in liquid nitrogen. Theharvested tissue is then stored at −80° C. until RNA preparation.

The CMz045 (Lib3076) cDNA library is generated from maize immature earmegaspore tissue. Seeds are planted at a depth of approximately 3 cminto 2-3 inch peat pots containing Metro 200 growing medium. After 2-3weeks growth they are transplanted into 10 inch pots containing the samegrowing medium. Plants are watered daily before transplantation andthree times a week after transplantation. Peters 15-16-17 fertilizer isapplied three times per week after transplanting at a strength of 150ppm N. Two to three times during the lifetime of the plant, fromtransplanting to flowering, a total of 900 mg Fe is added to each pot.Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles.The daytime temperature is approximately 80° F. and the nighttimetemperature is approximately 70° F. Supplemental lighting is provided by1000 W sodium vapor lamps. Tissue is collected from immature ear(megaspore) obtained from 7 week old plants. The immature ears areharvested from the 7 week old plants and are approximately 2.5 to 3 cmin length. The kernels are removed from the cob immediately frozen inliquid nitrogen. The harvested tissue is then stored at −80° C. untilRNA preparation.

The CMz047 (Lib3078) cDNA library is generated from maize CO₂ treatedhigh-exposure shoot tissue at the V10+ plant development stage. RX601maize seeds are sterilized for i minute with a 10% clorox solution. Theseeds are rolled in germination paper, and germinated in 0.5 mM calciumsulfate solution for two days ate 30° C. The seedlings are planted at adepth of approximately 3 cm into 2-3 inch peat pots containing Metro 200growing medium at a rate of 2-3 seedlings per pot. Twenty pots areplaced into a high CO₂ environment (approximately 1000 ppm CO₂). Twentyplants were grown under ambient greenhouse CO₂ (approximately 450 ppmCO₂). Plants are watered daily before transplantation and three times aweek after transplantation. Peters 20-20-20 fertilizer is also lightlyapplied. Maize plants are grown in the greenhouse in 15 hr day/9 hrnight cycles. The daytime temperature is approximately 80° F. and thenighttime temperature is approximately 70° F. Supplemental lighting isprovided by 1000 W sodium vapor lamps. At ten days post planting, theshoots from both atmosphere are frozen in liquid nitrogen and lightlyground. The roots are washed in deionized water to remove the supportmedia and the tissue is immediately transferred to liquid nitrogencontainers. The harvested tissue is then stored at −80° C. until RNApreparation.

The CMz048 (Lib3079) cDNA library is generated from maize basalendosperm transfer layer tissue at the V10+ plant development stage.Seeds are planted at a depth of approximately 3 cm into 2-3 inch peatpots containing Metro 200 growing medium. After 2-3 weeks growth theyare transplanted into 10 inch pots containing the same growing medium.Plants are watered daily before transplantation and three times a weekafter transplantation. Peters 15-16-17 fertilizer is applied three timesper week after transplanting at a strength of 150 ppm N. Two to threetimes during the lifetime of the plant, from transplanting to flowering,a total of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected from V10+ maize plants. The ear shoots, which areready for fertilization, are enclosed in a paper bag prior to silkemergence, to withhold the pollen. Kernels are harvested at 12 dayspost-pollination and placed on wet ice for dissection. The kernels arecross sectioned laterally, dissecting just above the pedicel region,including 1-2 mm of the lower endosperm and the basal endosperm transferregion. The pedicel and lower endosperm region containing the basalendosperm transfer layer is pooled and immediately frozen in liquidnitrogen. The harvested tissue is then stored at −80° C. until RNApreparation.

The CMz049 (Lib3088) cDNA library is generated from maize immatureanther tissue at the 7 week old immature tassel stage. Seeds are plantedat a depth of approximately 3 cm into 2-3 inch peat pots containingMetro 200 growing medium. After 2-3 weeks growth they are transplantedinto 10 inch pots containing the same growing medium. Plants are watereddaily before transplantation and three times a week aftertransplantation. Peters 15-16-17 fertilizer is applied three times perweek after transplanting at a strength of 150 ppm N. Two to three timesduring the lifetime of the plant, from transplanting to flowering, atotal of 900 mg Fe is added to each pot. Maize plants are grown in thegreenhouse in 15 hr day/9 hr night cycles. The daytime temperature isapproximately 80° F. and the nighttime temperature is approximately 70°F. Supplemental lighting is provided by 1000 W sodium vapor lamps.Tissue is collected when the maize plant is at the 7 week old immaturetassel stage. At this stage, prior to anthesis, the immature anthers aregreen and enclosed in the staminate spikelet. The developing anthers aredissected away from the 7 week old immature tassel and immediatelytransferred to liquid nitrogen container. The harvested tissue is thenstored at −80° C. until RNA preparation.

The CMz050 (Lib3114) cDNA library is generated from maize silk tissue atthe V10+ plant development stage. Seeds are planted at a depth ofapproximately 3 cm into 2-3 inch peat pots containing Metro 200 growingmedium. After 2-3 weeks growth they are transplanted into 10 inch potscontaining the same growing medium. Plants are watered daily beforetransplantation and three times a week after transplantation. Peters15-16-17 fertilizer is applied three times per week after transplantingat a strength of 150 ppm N. Two to three times during the lifetime ofthe plant, from transplanting to flowering, a total of 900 mg Fe isadded to each pot. Maize plants are grown in the greenhouse in 15 hrday/9 hr night cycles. The daytime temperature is approximately 80° F.and the nighttime temperature is approximately 70° F. Supplementallighting is provided by 1000 W sodium vapor lamps. Tissue is collectedwhen the maize plant is beyond the 10-leaf development stage and the earshoots are approximately 15-20 cm in length. The ears are pulled andsilks are separated from the ears and immediately transferred to liquidnitrogen containers. The harvested tissue is then stored at −80° C.until RNA preparation.

The SOYMON001 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) total leaf tissue atthe V4 plant development stage. Leaf tissue from 38, field grown V4stage plants is harvested from the 4^(th) node. Leaf tissue is removedfrom the plants and immediately frozen in dry-ice. The harvested tissueis then stored at −80° C. until RNA preparation.

The SOYMON002 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue at theV4 plant development stage. Root tissue from 76, field grown V4 stageplants is harvested. The root systems is cut from the soybean plant andwashed with water to free it from the soil and immediately frozen indry-ice. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON003 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotylaxis tissue harvested 2 day post-imbibition. Seeds are planted at adepth of approximately 2 cm into 2-3 inch peat pots containing Metromix350 medium. Trays are placed in an environmental chamber and grown at 12hr daytime/12 hr nighttime cycles. The daytime temperature isapproximately 29° C. and the nighttime temperature approximately 24° C.Soil is checked and watered daily to maintain even moisture conditions.Tissue is collected 2 days after the start of imbibition. The 2 daysafter imbibition samples are separated into 3 collections after removalof any adhering seed coat. At the 2 day stage, the hypocotyl axis isemerging from the soil. A few seedlings have cracked the soil surfaceand exhibited slight greening of the exposed cotyledons. The seedlingsare washed in water to remove soil, hypocotyl axis harvested andimmediately frozen in liquid nitrogen. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SOYMON004 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledontissue harvested 2 day post-imbibition. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium. Trays are placed in an environmental chamber and grown at 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions. Tissueis collected 2 days after the start of imbibition. The 2 days afterimbibition samples are separated into 3 collections after removal of anyadhering seed coat. At the 2 day stage, the hypocotyl axis is emergingfrom the soil. A few seedlings have cracked the soil surface andexhibited slight greening of the exposed cotyledons. The seedlings arewashed in water to remove soil, hypocotyl axis harvested and immediatelyfrozen in liquid nitrogen. The harvested tissue is then stored at −80°C. until RNA preparation.

The SOYMON005 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotylaxis tissue harvested 6 hour post-imbibition. Seeds are planted at adepth of approximately 2 cm into 2-3 inch peat pots containing Metromix350 medium. Trays are placed in an environmental chamber and grown at 12hr daytime/12 hr nighttime cycles. The daytime temperature isapproximately 29° C. and the nighttime temperature approximately 24° C.Soil is checked and watered daily to maintain even moisture conditions.Tissue is collected 6 hours after the start of imbibition. The 6 hoursafter imbibition samples are separated into 3 collections after removalof any adhering seed coat. The 6 hours after imbibition sample iscollected over the course of approximately 2 hours starting at 6 hourspost imbibition. At the 6 hours after imbibition stage, not allcotyledons have become fully hydrated and germination, or radicleprotrusion, has not occurred. The seedlings are washed in water toremove soil, hypocotyl axis harvested and immediately frozen in liquidnitrogen. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON006 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledonstissue harvest 6 hour post-imbibition. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium. Trays are placed in an environmental chamber and grown at 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions. Tissueis collected 6 hours after imbibition. The 6 hours after imbibitionsamples are separated into 3 collections after removal of any adheringseed coat. The 6 hours after imbibition sample is collected over thecourse of approximately 2 hours starting at 6 hours post-imbibition. Atthe 6 hours after imbibition, not all cotyledons have become fullyhydrated and germination or radicle protrusion, have not occurred. Theseedlings are washed in water to remove soil, cotyledon harvested andimmediately frozen in liquid nitrogen. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SOYMON007 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissueharvested 25 and 35 days post-flowering. Seed pods from field grownplants are harvested 25 and 35 days after flowering and the seedsextracted from the pods. Approximately 4.4 g and 19.3 g of seeds areharvested from the respective seed pods and immediately frozen in dryice. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON008 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissueharvested from 25 and 35 days post-flowering plants. Total leaf tissueis harvested from field grown plants. Approximately 19 g and 29 g ofleaves are harvested from the fourth node of the plant 25 and 35 dayspost-flowering and immediately frozen in dry ice. The harvested tissueis then stored at −80° C. until RNA preparation.

The SOYMON009 cDNA library is generated from soybean cutlivar C1944(USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) pod and seedtissue harvested 15 days post-flowering. Pods from field grown plantsare harvested 15 days post-flowering. Approximately 3 g of pod tissue isharvested and immediately frozen in dry-ice. The harvested tissue isthen stored at −80° C. until RNA preparation.

The SOYMON010 cDNA library is generated from soybean cultivar C1944(USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) seed tissueharvested 40 days post-flowering. Pods from field grown plants areharvested 40 days post-flowering. Pods and seeds are separated,approximately 19 g of seed tissue is harvested and immediately frozen indry-ice. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON011 cDNA library is generated from soybean cultivarsCristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) andFT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves areharvested from plants grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions.Approximately 30 g of leaves are harvested from the 4′ node of each ofthe Cristalina and FT108 cultivars and immediately frozen in dry ice.The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON012 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue. Leavesfrom field grown plants are harvested from the fourth node 15 dayspost-flowering. Approximately 12 g of leaves are harvested andimmediately frozen in dry ice. The harvested tissue is then stored at−80° C. until RNA preparation.

The SOYMON013 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root and noduletissue. Approximately, 28 g of root tissue from field grown plants isharvested 15 days post-flowering. The root system is cut from thesoybean plant, washed with water to free it from the soil andimmediately frozen in dry-ice. The harvested tissue is then stored at−80° C. until RNA preparation.

The SOYMON014 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissueharvested 25 and 35 days after flowering. Seed pods from field grownplants are harvested 15 days after flowering and the seeds extractedfrom the pods. Approximately 5 g of seeds are harvested from therespective seed pods and immediately frozen in dry ice. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SOYMON015 cDNA is generated from soybean cultivar Asgrow 3244(Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 45and 55 days post-flowering. Seed pods from field grown plants areharvested 45 and 55 days after flowering and the seeds extracted fromthe pods. Approximately 19 g and 31 g of seeds are harvested from therespective seed pods and immediately frozen in dry ice. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SOYMON016 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue.Approximately, 61 g and 38 g of root tissue from field grown plants isharvested 25 and 35 days post-flowering is harvested. The root system iscut from the soybean plant and washed with water to free it from thesoil. The tissue is placed in 14 ml polystyrene tubes and immediatelyfrozen in dry-ice. The harvested tissue is then stored at −80° C. untilRNA preparation.

The SOYMON017 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue.Approximately 28 g of root tissue from field grown plants is harvested45 and 55 days post-flowering. The root system is cut from the soybeanplant, washed with water to free it from the soil and immediately frozenin dry-ice. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON018 cDNA is generated from soybean cultivar Asgrow 3244(Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 45and 55 days post-flowering. Leaves from field grown plants are harvested45 and 55 days after flowering from the fourth node. Approximately 27 gand 33 g of seeds are harvested from the respective seed pods andimmediately frozen in dry ice. The harvested tissue is then stored at−80° C. until RNA preparation.

The SOYMON019 cDNA library is generated from soybean cultivarsCristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) andFT108 (Monsoy, Brazil) (tropical germ plasma) root tissue. Roots areharvested from plants grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions.Approximately 50 g and 56 g of roots are harvested from each of theCristalina and FT108 cultivars and immediately frozen in dry ice. Theharvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON020 cDNA is generated from soybean cultivar Asgrow 3244(Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 65and 75 days post-flowering. Seed pods from field grown plants areharvested 45 and 55 days after flowering and the seeds extracted fromthe pods. Approximately 14 g and 31 g of seeds are harvested from therespective seed pods and immediately frozen in dry ice. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SOYMON021 cDNA library is generated from Soybean CystNematode-resistant soybean cultivar Hartwig (USDA Soybean GermplasmCollection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in tissueculture at room temperature. At approximately 6 weeks post-germination,the plants are exposed to sterilized Soybean Cyst Nematode eggs.Infection is then allowed to progress for 10 days. After the 10 dayinfection process, the tissue is harvested. Agar from the culture mediumand nematodes are removed and the root tissue is immediately frozen indry ice. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON022 (Lib3030) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) partiallyopened flower tissue. Partially to fully opened flower tissue isharvested from plants grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions. A totalof 3 g of flower tissue is harvested and immediately frozen in dry ice.The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON023 cDNA library is generated from soybean genotype BW211SNull (Tohoku University, Morioka, Japan) seed tissue harvested 15 and 40days post-flowering. Seed pods from field grown plants are harvested 15and 40 days post-flowering and the seeds extracted from the pods.Approximately 0.7 g and 14.2 g of seeds are harvested from therespective seed pods and immediately frozen in dry ice. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SOYMON024 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) internode-2 tissueharvested 18 days post-imbibition. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium. The plants are grown in a greenhouse for 18 days after the startof imbibition at ambient temperature. Soil is checked and watered dailyto maintain even moisture conditions. Stem tissue is harvested 18 daysafter the start of imbibition. The samples are divided into hypocotyland internodes 1 through 5. The fifth internode contains some leaf budmaterial. Approximately 3 g of each sample is harvested and immediatelyfrozen in dry ice. The harvested tissue is then stored at −80° C. untilRNA preparation.

The SOYMON025 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissueharvested 65 days post-flowering. Leaves are harvested from the fourthnode of field grown plants 65 days post-flowering. Approximately 18.4 gof leaf tissue is harvested and immediately frozen in dry ice. Theharvested tissue is then stored at −80° C. until RNA preparation.

SOYMON026 cDNA library is generated from soybean cultivar Asgrow 3244(Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue harvested 65and 75 days post-flowering. Approximately 27 g and 40 g of root tissuefrom field grown plants is harvested 65 and 75 days post-flowering. Theroot system is cut from the soybean plant, washed with water to free itfrom the soil and immediately frozen in dry-ice. The harvested tissue isthen stored at −80° C. until RNA preparation.

The SOYMON027 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissueharvested 25 days post-flowering. Seed pods from field grown plants areharvested 25 days post-flowering and the seeds extracted from the pods.Approximately 17 g of seeds are harvested from the seed pods andimmediately frozen in dry ice. The harvested tissue is then stored at−80° C. until RNA preparation.

The SOYMON028 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressedroot tissue. The plants are grown in an environmental chamber under 12hr daytime/12 hr nighttime cycles. The daytime temperature isapproximately 29° C. and the nighttime temperature 24° C. Soil ischecked and watered daily to maintain even moisture conditions. At theR3 stage of development, water is withheld from half of the plantcollection (drought stressed population). After 3 days, half of theplants from the drought stressed condition and half of the plants fromthe control population are harvested. After another 3 days (6 days postdrought induction) the remaining plants are harvested. A total of 27 gand 40 g of root tissue is harvested and immediately frozen in dry ice.The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON029 cDNA library is generated from Soybean CystNematode-resistant soybean cultivar PI07354 (USDA Soybean GermplasmCollection, Urbana, Ill. U.S.A.) root tissue. Late fall to early wintergreenhouse grown plants are exposed to Soybean Cyst Nematode eggs. At 10days post-infection, the plants are uprooted, rinsed briefly and theroots frozen in liquid nitrogen. Approximately 20 grams of root tissueis harvested from the infected plants. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SOYMON030 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) flower bud tissue.Seeds are planted at a depth of approximately 2 cm into 2-3 inch peatpots containing Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 29° C. and the nighttimetemperature approximately 24° C. Soil is checked and watered daily tomaintain even moisture conditions. Flower buds are removed from theplant at the pedicel. A total of 100 mg of flower buds are harvested andimmediately frozen in liquid nitrogen. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SOYMON031 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) carpel and stamentissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inchpeat pots containing Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 29° C. and the nighttimetemperature approximately 24° C. Soil is checked and watered daily tomaintain even moisture conditions. Flower buds are removed from theplant at the pedicel. Flowers are dissected to separate petals, sepalsand reproductive structures (carpels and stamens). A total of 300 mg ofcarpel and stamen tissue are harvested and immediately frozen in liquidnitrogen. The harvested tissue is then stored at −80° C. until RNApreparation.

The SOYMON032 cDNA library is prepared from the Asgrow cultivar A4922(Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry soybeanseed meristem tissue. Surface sterilized seeds are germinated in liquidmedia for 24 hours. The seed axis is then excised from the barelygerminating seed, placed on tissue culture media and incubated overnightat 20° C. in the dark. The supportive tissue is removed from the explantprior to harvest. Approximately 570 mg of tissue is harvested and frozenin liquid nitrogen. The harvested tissue is then stored at −80° C. untilRNA preparation.

The SOYMON033 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heat-shockedseedling tissue without cotyledons. Seeds are imbibed and germinated invermiculite for 2 days under constant illumination. After 48 hours, theseedlings are transferred to an incubator set at 40° C. under constantillumination. After 30, 60 and 180 minutes seedlings are harvested anddissected. A portion of the seedling consisting of the root, hypocotyland apical hook is frozen in liquid nitrogen and stored at −80° C. Theseedlings after 2 days of imbibition are beginning to emerge from thevermiculite surface. The apical hooks are dark green in appearance.Total RNA and poly A⁺ RNA is prepared from equal amounts of pooledtissue.

The SOYMON034 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) cold-shockedseedling tissue without cotyledons. Seeds are imbibed and germinated invermiculite for 2 days under constant illumination. After 48 hours, theseedlings are transferred to a cold room set at 5° C. under constantillumination. After 30, 60 and 180 minutes seedlings are harvested anddissected. A portion of the seedling consisting of the root, hypocotyland apical hook is frozen in liquid nitrogen and stored at −80° C. Theseedlings after 2 days of imbibition are beginning to emerge from thevermiculite surface. The apical hooks are dark green in appearance.

The SOYMON035 cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed coat tissue.Seeds are planted at a depth of approximately 2 cm into 2-3 inch peatpots containing Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 29° C. and the nighttimetemperature 24° C. Soil is checked and watered daily to maintain evenmoisture conditions. Seeds are harvested from mid to nearly fullmaturation (seed coats are not yellowing). The entire embryo proper isremoved from the seed coat sample and the seed coat tissue are harvestedand immediately frozen in liquid nitrogen. The harvested tissue is thenstored at −80° C. until RNA preparation.

The SOYMON036 cDNA library is generated from soybean cultivars PI171451,PI227687 and PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill.U.S.A.) insect challenged leaves. Plants from each of the threecultivars are grown in screenhouse conditions. The screenhouse isdivided in half and one half of the screenhouse is infested with soybeanlooper and the other half infested with velvetbean caterpillar. A singleleaf is taken from each of the representative plants at 3 different timepoints, 11 days after infestation, 2 weeks after infestation and 5 weeksafter infestation and immediately frozen in liquid nitrogen. Theharvested tissue is then stored at −80° C. until RNA preparation. TotalRNA and poly A+ RNA is isolated from pooled tissue consisting of equalquantities of all 18 samples (3 genotypes×3 sample times×2 insectgenotypes).

The SOYMON037 cDNA library is generated from soybean cultivar A3244(Asgrow Seed Company, Des Moines, Iowa U.S.A.) etiolated axis andradical tissue. Seeds are planted in moist vermiculite, wrapped and keptat room temperature in complete darkness until harvest. Etiolated axisand hypocotyl tissue is harvested at 2, 3 and 4 days post-planting. Atotal of 1 gram of each tissue type is harvested at 2, 3 and 4 daysafter planting and immediately frozen in liquid nitrogen. The harvestedtissue is then stored at −80° C. until RNA preparation.

The SOYMON038 cDNA library is generated from soybean variety AsgrowA3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dryseeds. Explants are prepared for transformation after germination ofsurface-sterilized seeds on solid tissue media. After 6 days, at 28° C.and 18 hours of light per day, the germinated seeds are cold shocked at4° C. for 24 hours. Meristemic tissue and part of the hypocotyl isremove and cotyledon excised. The prepared explant is then wounded forAgrobacterium infection. The 2 grams of harvested tissue is frozen inliquid nitrogen and stored at −80° C. until RNA preparation.

The Soy51 (LIB3027) cDNA library is prepared from equal amounts tissueharvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue.Single stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio) and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. The non-hybridizedsingle stranded molecules remaining after hybrid capture are convertedto double stranded form and represent the primary normalized library.

The Soy52 (LIB3028) cDNA library is generated from normalized flowerDNA. Single stranded DNA representing approximately 1×10⁶ colony formingunits of SOYMON022 harvested tissue is used as the starting material fornormalization. RNA, complementary to the single stranded DNA, issynthesized using the double stranded DNA as a template. BiotinylateddATP is incorporated into the RNA during the synthesis reaction. Thesingle stranded DNA is mixed with the biotinylated RNA in a 1:10 molarratio) and allowed to hybridize. DNA-RNA hybrids are captured onDynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success,N.Y. U.S.A.). The dynabeads with captured hybrids are collected with amagnet. The non-hybridized single stranded molecules remaining afterhybrid capture are converted to double stranded form and represent theprimary normalized library.

The Soy53 (LIB3039) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedlingshoot apical meristem tissue. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium and the plants are grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature 24° C. Soil is checked and watereddaily to maintain even moisture conditions. Apical tissue is harvestedfrom seedling shoot meristem tissue, 7-8 days after the start ofimbibition. The apex of each seedling is dissected to include the fifthnode to the apical meristem. The fifth node corresponds to the thirdtrifoliate leaf in the very early stages of development. Stipulescompletely envelop the leaf primordia at this time. A total of 200 mg ofapical tissue is harvested and immediately frozen in liquid nitrogen.The harvested tissue is then stored at −80° C. until RNA preparation.

The Soy54 (LIB3040) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heart totorpedo stage embryo tissue. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium and the plants are grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature 24° C. Soil is checked and watereddaily to maintain even moisture conditions. Seeds are collected andembryos removed from surrounding endosperm and maternal tissues. Embryosfrom globular to young torpedo stages (by corresponding analogy toArabidopsis) are collected with a bias towards the middle of thisspectrum. Embryos which are beginning to show asymmetric development ofcotyledons are considered the upper developmental boundary for thecollection and are excluded. A total of 12 mg embryo tissue is frozen inliquid nitrogen. The harvested tissue is stored at −80° C. until RNApreparation.

Soy55 (LIB3049) cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) young seed tissue.Seeds are planted at a depth of approximately 2 cm into 2-3 inch peatpots containing Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 29° C. and the nighttimetemperature 24° C. Soil is checked and watered daily to maintain evenmoisture conditions. Seeds are collected from very young pods (5 to 15days after flowering). A total of 100 mg of seeds are harvested andfrozen in liquid nitrogen. The harvested tissue is stored at −80° C.until RNA preparation.

Soy56 (LIB3029) cDNA library is prepared from equal amounts tissueharvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue.Single stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. The non-hybridizedsingle stranded molecules remaining after hybrid capture are notconverted to double stranded form and represent a non-normalized seedpool for comparison to Soy51 cDNA libraries.

The Soy58 (LIB3050) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) droughtstressed root tissue subtracted from control root tissue. Seeds areplanted at a depth of approximately 2 cm into 2-3 inch peat potscontaining Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 29° C. and the nighttimetemperature 24° C. Soil is checked and watered daily to maintain evenmoisture conditions. At the R3 stage of the plant drought is induced bywithholding water. After 3 and 6 days root tissue from both droughtstressed and control (watered regularly) plants are collected and frozenin dry-ice. The harvested tissue is stored at −80° C. until RNApreparation. For subtraction, target cDNA is made from the droughtstressed tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy59 (LIB3051) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) endospermtissue. Seeds are germinated on paper towels under laboratory ambientlight conditions. At 8, 10 and 14 hours after imbibition, the seed coatsare harvested. The endosperm consists of a very thin layer of tissueaffixed to the inside of the seed coat. The seed coat and endosperm arefrozen immediately after harvest in liquid nitrogen. The harvestedtissue is stored at −80° C. until RNA preparation.

The Soy60 (LIB3072) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) droughtstressed seed plus pod subtracted from control seed plus pod tissue.Seeds are planted at a depth of approximately 2 cm into 2-3 inch peatpots containing Metromix 350 medium and the plants are grown in anenvironmental chamber under 12 hr daytime/12 hr nighttime cycles. Thedaytime temperature is approximately 26° C. and the nighttimetemperature 21° C. and 70% relative humidity. Soil is checked andwatered daily to maintain even moisture conditions. At the R3 stage ofthe plant drought is induced by withholding water. After 3 and 6 daysseeds and pods from both drought stressed and control (wateredregularly) plants are collected from the fifth and sixth node and frozenin dry-ice. The harvested tissue is stored at −80° C. until RNApreparation. For subtraction, target cDNA is made from the droughtstressed tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy61 (LIB3073) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acidtreated seedling subtracted from control tissue. Seeds are planted at adepth of approximately 2 cm into 2-3 inch peat pots containing Metromix350 medium and the plants are grown in a greenhouse. The daytimetemperature is approximately 29.4° C. and the nighttime temperature 20°C. Soil is checked and watered daily to maintain even moistureconditions. At 9 days post planting, the plantlets are sprayed witheither control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500,Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants aresprayed until runoff and the soil and the stem is socked with thespraying solution. At 18 hours post application of jasmonic acid, thesoybean plantlets appear growth retarded. After 18 hours, 24 hours and48 hours post treatment, the cotyledons are removed and the remainingleaf and stem tissue above the soil is harvested and frozen in liquidnitrogen. The harvested tissue is stored at −80° C. until RNApreparation. To make RNA, the three sample timepoints were combined andground. For subtraction, target cDNA is made from the jasmonic acidtreated tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library'sconstruction, the eighth fraction of the cDNA size fractionation stepwas used for ligation.

The Soy62 (LIB3074) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acidtreated seedling subtracted from control tissue. Seeds are planted at adepth of approximately 2 cm into 2-3 inch peat pots containing Metromix350 medium and the plants are grown in a greenhouse. The daytimetemperature is approximately 29.4° C. and the nighttime temperature 20°C. Soil is checked and watered daily to maintain even moistureconditions. At 9 days post planting, the plantlets are sprayed witheither control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500,Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants aresprayed until runoff and the soil and the stem is socked with thespraying solution. At 18 hours post application of jasmonic acid, thesoybean plantlets appear growth retarded. After 18 hours, 24 hours and48 hours post treatment, the cotyledons are removed and the remainingleaf and stem tissue above the soil is harvested and frozen in liquidnitrogen. The harvested tissue is stored at −80° C. until RNApreparation. To make RNA, the three sample timepoints were combined andground. For subtraction, target cDNA is made from the jasmonic acidtreated tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library'sconstruction, the ninth fraction of the cDNA size fractionation step wasused for ligation.

The Soy65 (LIB3107) 07cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.)drought-stressed abscission zone tissue. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium and the plants are grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature 24° C. Soil is checked and watereddaily to maintain even moisture conditions. Plants are irrigated with15-16-17 Peter's Mix. At the R3 stage of development, drought is imposedby withholding water. At 3, 4, 5 and 6 days, tissue is harvested andwilting is not obvious until the fourth day. Abscission layers fromreproductive organs are harvested by cutting less than one millimeterproximal and distal to the layer and immediately frozen in liquidnitrogen. The harvested tissue is stored at −80° C. until RNApreparation.

The Soy66 (LIB3109) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) non-droughtstressed abscission zone tissue. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium and the plants are grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions. Plantsare irrigated with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, controlabscission layer tissue is harvested. Abscission layers fromreproductive organs are harvested by cutting less than one millimeterproximal and distal to the layer and immediately frozen in liquidnitrogen. The harvested tissue is stored at −80° C. until RNApreparation.

Soy67 (LIB3065) cDNA library is prepared from equal amounts tissueharvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue.Single stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio) and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. Captured hybrids areeluted with water.

Soy68 (LIB3052) cDNA library is prepared from equal amounts tissueharvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue.Single stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio) and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. Captured hybrids areeluted with water.

Soy69 (LIB3053) cDNA library is generated from soybean cultivarsCristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) andFT108 (Monsoy, Brazil) (tropical germ plasma) normalized leaf tissue.Leaves are harvested from plants grown in an environmental chamber under12 hr daytime/12 hr nighttime cycles. The daytime temperature isapproximately 29° C. and the nighttime temperature approximately 24° C.Soil is checked and watered daily to maintain even moisture conditions.Approximately 30 g of leaves are harvested from the 4′ node of each ofthe Cristalina and FT108 cultivars and immediately frozen in dry ice.The harvested tissue is then stored at −80° C. until RNA preparation.Single stranded and double stranded DNA representing approximately 1×10⁶colony forming units are isolated using standard protocols. RNA,complementary to the single stranded DNA, is synthesized using thedouble stranded DNA as a template. Biotinylated dATP is incorporatedinto the RNA during the synthesis reaction. The single stranded DNA ismixed with the biotinylated RNA in a 1:10 molar ratio) and allowed tohybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.). The dynabeadswith captured hybrids are collected with a magnet. The non-hybridizedsingle stranded molecules remaining after hybrid capture are convertedto double stranded form and represent the primary normalized library.

Soy70 (LIB3055) cDNA library is generated from soybean cultivarsCristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) andFT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves areharvested from plants grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions.Approximately 30 g of leaves are harvested from the 4^(th) node of eachof the Cristalina and FT108 cultivars and immediately frozen in dry ice.The harvested tissue is then stored at −80° C. until RNA preparation.

Soy71 (LIB3056) cDNA library is generated from soybean cultivarsCristalina and FT108 (tropical germ plasma) root tissue. Roots areharvested from plants grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately29° C. and the nighttime temperature approximately 24° C. Soil ischecked and watered daily to maintain even moisture conditions.Approximately 50 g and 56 g of roots are harvested from each of theCristalina and FT108 cultivars and immediately frozen in dry ice. Theharvested tissue is then stored at −80° C. until RNA preparation.

Soy72 (LIB3093) cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressedleaf control tissue. Seeds are planted at a depth of approximately 2 cminto 2-3 inch peat pots containing Metromix 350 medium and the plantsare grown in an environmental chamber under 12 hr daytime/12 hrnighttime cycles. The daytime temperature is approximately 26° C. andthe nighttime temperature 21° C. and 70% relative humidity. Soil ischecked and watered daily to maintain even moisture conditions. At theR3 stage of the plant drought is induced by withholding water. After 3and 6 days seeds and pods from both drought stressed and control(watered regularly) plants are collected from the fifth and sixth nodeand frozen in dry-ice. The harvested tissue is stored at −80° C. untilRNA preparation. For subtraction, target cDNA is made from the droughtstressed tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

Soy73 (LIB3093) cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressedleaf subtracted from control tissue. Seeds are planted at a depth ofapproximately 2 cm into 2-3 inch peat pots containing Metromix 350medium and the plants are grown in an environmental chamber under 12 hrdaytime/12 hr nighttime cycles. The daytime temperature is approximately26° C. and the nighttime temperature 21° C. and 70% relative humidity.Soil is checked and watered daily to maintain even moisture conditions.At the R3 stage of the plant drought is induced by withholding water.After 3 and 6 days seeds and pods from both drought stressed and control(watered regularly) plants are collected from the fifth and sixth nodeand frozen in dry-ice. The harvested tissue is stored at −80° C. untilRNA preparation. For subtraction, target cDNA is made from the droughtstressed tissue total RNA using the SMART cDNA synthesis system fromClonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driverfirst strand cDNA is covalently linked to Dynabeads following a protocolsimilar to that described in the Dynal literature (Dynabeads, DynalCorporation, Lake Success, N.Y. U.S.A.). The target cDNA is then heatdenatured and the second strand trapped using Dynabeads oligo-dT. Thetarget second strand cDNA is then hybridized to the driver cDNA in 400μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. Aftereach hybridization, the hybridization solution is removed from thesystem and the hybridized target cDNA removed from the driver by heatdenaturation in water. After hybridization, the remaining cDNA istrapped with Dynabeads oligo-dT. The trapped cDNA is then amplified asin previous PCR based libraries and the resulting cDNA ligated into thepSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy76 (Lib3106) cDNA library is generated from soybean cultivarAsgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acidand arachidonic treated seedling subtracted from control tissue. Seedsare planted at a depth of approximately 2 cm into 2-3 inch peat potscontaining Metromix 350 medium and the plants are grown in a greenhouse.The daytime temperature is approximately 29.4° C. and the nighttimetemperature 20° C. Soil is checked and watered daily to maintain evenmoisture conditions. At 9 days post planting, the plantlets are sprayedwith either control buffer of 0.1% Tween-20 or jasmonic acid (SigmaJ-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20.Plants are sprayed until runoff and the soil and the stem is socked withthe spraying solution. At 18 hours post application of jasmonic acid,the soybean plantlets appear growth retarded. Arachidonic treatedseedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20.After 18 hours, 24 hours and 48 hours post treatment, the cotyledons areremoved and the remaining leaf and stem tissue above the soil isharvested and frozen in liquid nitrogen. The harvested tissue is storedat −80° C. until RNA preparation. To make RNA, the three sampletimepoints were combined and ground. The RNA from the arachidonictreated seedlings is isolated separately. For subtraction, target cDNAis made from the jasmonic acid treated tissue total RNA using the SMARTcDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto,Calif. U.S.A.). Driver first strand cDNA is covalently linked toDynabeads following a protocol similar to that described in the Dynalliterature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).The target cDNA is then heat denatured and the second strand trappedusing Dynabeads oligo-dT. The target second strand cDNA is thenhybridized to the driver cDNA in 400 μl 2×SSPE for two rounds ofhybridization at 65° C. and 20 hours. After each hybridization, thehybridization solution is removed from the system and the hybridizedtarget cDNA removed from the driver by heat denaturation in water. Afterhybridization, the remaining cDNA is trapped with Dynabeads oligo-dT.The trapped cDNA is then amplified as in previous PCR based librariesand the resulting cDNA ligated into the pSPORT vector (Invitrogen,Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA isligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.) inorder to capture some of the smaller transcripts characteristic ofantifungal proteins.

Soy77 (LIB3108) cDNA library is generated from soybean cultivar Asgrow3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acidcontrol tissue. Seeds are planted at a depth of approximately 2 cm into2-3 inch peat pots containing Metromix 350 medium and the plants aregrown in a greenhouse. The daytime temperature is approximately 29.4° C.and the nighttime temperature 20° C. Soil is checked and watered dailyto maintain even moisture conditions. At 9 days post planting, theplantlets are sprayed with either control buffer of 0.1% Tween-20 orjasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in0.1% Tween-20. Plants are sprayed until runoff and the soil and the stemis socked with the spraying solution. At 18 hours post application ofjasmonic acid, the soybean plantlets appear growth retarded. Arachidonictreated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1%Tween-20. After 18 hours, 24 hours and 48 hours post treatment, thecotyledons are removed and the remaining leaf and stem tissue above thesoil is harvested and frozen in liquid nitrogen. The harvested tissue isstored at −80° C. until RNA preparation. To make RNA, the three sampletimepoints were combined and ground. The RNA from the arachidonictreated seedlings is isolated separately. For subtraction, target cDNAis made from the jasmonic acid treated tissue total RNA using the SMARTcDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto,Calif. U.S.A.). Driver first strand cDNA is covalently linked toDynabeads following a protocol similar to that described in the Dynalliterature (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).The target cDNA is then heat denatured and the second strand trappedusing Dynabeads oligo-dT. The target second strand cDNA is thenhybridized to the driver cDNA in 400 μl 2×SSPE for two rounds ofhybridization at 65° C. and 20 hours. After each hybridization, thehybridization solution is removed from the system and the hybridizedtarget cDNA removed from the driver by heat denaturation in water. Afterhybridization, the remaining cDNA is trapped with Dynabeads oligo-dT.The trapped cDNA is then amplified as in previous PCR based librariesand the resulting cDNA ligated into the pSPORT vector (Invitrogen,Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA isligated into the pSPORT vector in order to capture some of the smallertranscripts characteristic of antifungal proteins.

The stored RNA is purified using Trizol reagent from Life Technologies(Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially asrecommended by the manufacturer. Poly A+ RNA (mRNA) is purified usingmagnetic oligo dT beads essentially as recommended by the manufacturer(Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).

Construction of plant cDNA libraries is well-known in the art and anumber of cloning strategies exist. A number of cDNA libraryconstruction kits are commercially available. The Superscript™ PlasmidSystem for cDNA synthesis and Plasmid Cloning (Gibco BRL, LifeTechnologies, Gaithersburg, Md. U.S.A.) is used, following theconditions suggested by the manufacturer.

Normalized libraries are made using essentially the Soares procedure(Soares et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:9228-9232 (1994), theentirety of which is herein incorporated by reference). This approach isdesigned to reduce the initial 10,000-fold variation in individual cDNAfrequencies to achieve abundances within one order of magnitude whilemaintaining the overall sequence complexity of the library. In thenormalization process, the prevalence of high-abundance cDNA clonesdecreases dramatically, clones with mid-level abundance are relativelyunaffected and clones for rare transcripts are effectively increased inabundance.

EXAMPLE 2

The cDNA libraries are plated on LB agar containing the appropriateantibiotics for selection and incubated at 37° for a sufficient time toallow the growth of individual colonies. Single colonies areindividually placed in each well of a 96-well microtiter platescontaining LB liquid including the selective antibiotics. The plates areincubated overnight at approximately 37° C. with gentle shaking topromote growth of the cultures. The plasmid DNA is isolated from eachclone using Qiaprep plasmid isolation kits, using the conditionsrecommended by the manufacturer (Qiagen Inc., Santa Clara, Calif.U.S.A.).

Template plasmid DNA clones are used for subsequent sequencing. Forsequencing, the ABI PRISM dRhodamine Terminator Cycle Sequencing ReadyReaction Kit with AmpliTaq® DNA Polymerase, FS, is used (PE AppliedBiosystems, Foster City, Calif. U.S.A.).

EXAMPLE 3

Nucleic acid sequences that encode for the following proteins:methionine adenosyltransferase, S-adenosylmethionine decarboxylase,aspartate kinase, aspartate-semialdehyde dehydrogenase,O-succinylhomoserine (thiol)-lyase, cystathionine β-lyase,5-methyltetrahydropteroyltriglutamate, adenosylhomocysteinase,cystathionine β-synthase, cystathionine γ-lyase and O-acetylhomoserine(thiol)-lyase are identified from the Monsanto EST PhytoSeq databaseusing TBLASTN (default values) (TBLASTN compares a protein query againstthe six reading frames of a nucleic acid sequence). Matches found withBLAST P values equal or less than 0.001 (probability) or BLAST Score ofequal or greater than 90 are classified as hits. If the program used todetermine the hit is HMMSW then the score refers to HMMSW score.

In addition, the GenBank database is searched with BLASTN and BLASTX(default values) using ESTs as queries. EST that pass the hitprobability threshold of 10e⁻⁸ for the following enzymes are combinedwith the hits generated by using TBLASTN (described above) andclassified by enzyme (see Table A below).

A cluster refers to a set of overlapping clones in the PhytoSeqdatabase. Such an overlapping relationship among clones is designated asa “cluster” when BLAST scores from pairwise sequence comparisons of themember clones meets a predetermined minimum value or product score of 50or more (Product Score=(BLAST SCORE×Percentage Identity)/(5×minimum[length (Seq1), length (Seq2)]))

Since clusters are formed on the basis of single-linkage relationships,it is possible for two non-overlapping clones to be members of the samecluster if, for instance, they both overlap a third clone with at leastthe predetermined minimum BLAST score (stringency). A cluster ID isarbitrarily assigned to all of those clones which belong to the samecluster at a given stringency and a particular clone will belong to onlyone cluster at a given stringency. If a cluster contains only a singleclone (a “singleton”), then the cluster ID number will be negative, withan absolute value equal to the clone ID number of its single member.Clones grouped in a cluster in most cases represent a contiguoussequence. TABLE A* METHIONINE ADENOSYLTRANSFERASE (EC 2.5.1.6) Seq No.Cluster ID CloneID Library NCBI gi Method Score P-value % Ident 1-700019427 700019427H1 SATMON001 g17262 BLASTX 120 1e−11 92 2 -700074004700074004H1 SATMON007 g1778820 BLASTN 830 1e−60 80 3 -700149718700149718H1 SATMON007 g1778820 BLASTN 263 1e−13 80 4 -700220251700220251H1 SATMON011 g1127582 BLASTN 243 1e−9 90 5 -700458196700458196H1 SATMON029 g1778820 BLASTN 202 1e−22 88 6 -701172459701172459H1 SATMONN05 g882334 BLASTN 801 1e−57 89 7 -L1436317LIB143-062-Q1-E1-B6 LIB143 g1778820 BLASTN 341 1e−19 77 8 -L1893126LIB189-021-Q1-E1-F4 LIB189 g1778820 BLASTN 629 1e−41 88 9 -L1894542LIB189-033-Q1-E1-H12 LIB189 g1778820 BLASTN 564 1e−60 76 10 -L30622839LIB3062-028-Q1-K1-C2 LIB3062 g1778820 BLASTN 625 1e−65 74 11 -L30671966LIB3067-036-Q1-K1-D8 LIB3067 g1655576 BLASTX 118 1e−25 85 12 -L30682166LIB3068-004-Q1-K1-D5 LIB3068 g497900 BLASTX 111 1e−29 47 13 1LIB143-036-Q1-E1-G7 LIB143 g450548 BLASTN 1766 1e−138 88 14 1LIB3060-013-Q1-K1-F7 LIB3060 g1778820 BLASTN 1702 1e−133 90 15 1LIB143-013-Q1-E1-A6 LIB143 g1778820 BLASTN 1712 1e−133 88 16 1LIB148-007-Q1-E1-B8 LIB148 g450548 BLASTN 1693 1e−132 88 17 1LIB3079-003-Q1-K1-G1 LIB3079 g1778820 BLASTN 1236 1e−130 88 18 1LIB3066-011-Q1-K1-E4 LIB3066 g1778820 BLASTN 1661 1e−129 86 19 1LIB3068-008-Q1-K1-D2 LIB3068 g450548 BLASTN 1192 1e−128 88 20 1LIB143-061-Q1-E1-E7 LIB143 g1778820 BLASTN 1638 1e−127 88 21 1LIB189-006-Q1-E1-B2 LIB189 g1778820 BLASTN 1627 1e−126 86 22 1LIB143-031-Q1-E1-C7 LIB143 g450548 BLASTN 1595 1e−124 86 23 1LIB3068-011-Q1-K1-B12 LIB3068 g450548 BLASTN 1552 1e−123 87 24 1LIB143-013-Q1-E1-G11 LIB143 g450548 BLASTN 1585 1e−123 88 25 1LIB3062-057-Q1-K1-H9 LIB3062 g450548 BLASTN 1577 1e−122 89 26 1LIB143-008-Q1-E1-G9 LIB143 g450548 BLASTN 1556 1e−120 89 27 1LIB3068-058-Q1-K1-B7 LIB3068 g450548 BLASTN 1470 1e−118 86 28 1LIB3067-048-Q1-K1-G5 LIB3067 g1778820 BLASTN 1518 1e−117 89 29 1LIB143-049-Q1-E1-A12 LIB143 g1778820 BLASTN 1480 1e−114 91 30 1LIB3062-010-Q1-K1-C7 LIB3062 g1778820 BLASTN 1145 1e−113 87 31 1LIB3068-055-Q1-K1-E10 LIB3068 g450548 BLASTN 1451 1e−112 88 32 1LIB189-025-Q1-E1-H1 LIB189 g1778820 BLASTN 1460 1e−112 90 33 1LIB143-066-Q1-E1-F4 LIB143 g1778820 BLASTN 1309 1e−111 86 34 1LIB143-003-Q1-E1-D10 LIB143 g1778820 BLASTN 1440 1e−111 87 35 1LIB3062-033-Q1-K1-G1 LIB3062 g1778820 BLASTN 1449 1e−111 86 36 1LIB3066-029-Q1-K1-H11 LIB3066 g1778820 BLASTN 1427 1e−110 88 37 1LIB3068-016-Q1-K1-D9 LIB3068 g450548 BLASTN 1213 1e−109 87 38 1LIB3062-027-Q1-K1-B11 LIB3062 g1778820 BLASTN 1187 1e−108 87 39 1LIB3068-048-Q1-K1-G4 LIB3068 g1778820 BLASTN 1410 1e−108 82 40 1LIB148-020-Q1-E1-B6 LIB148 g1778820 BLASTN 1088 1e−107 84 41 1LIB3066-046-Q1-K1-F2 LIB3066 g1778820 BLASTN 1400 1e−107 90 42 1700084130H1 SATMON011 g1778820 BLASTN 960 1e−106 92 43 1 700092863H1SATMON008 g1778820 BLASTN 1388 1e−106 92 44 1 LIB3068-043-Q1-K1-A12LIB3068 g450548 BLASTN 827 1e−105 81 45 1 700092659H1 SATMON008 g1778820BLASTN 1359 1e−104 92 46 1 LIB143-038-Q1-E1-H11 LIB143 g450548 BLASTN1345 1e−103 84 47 1 LIB3062-028-Q1-K1-F12 LIB3062 g1778820 BLASTN 10821e−102 82 48 1 700103135H1 SATMON010 g1778820 BLASTN 1306 1e−100 93 49 1LIB3078-050-Q1-K1-D9 LIB3078 g1778820 BLASTN 1309 1e−100 86 50 1700084823H1 SATMON011 g1778820 BLASTN 1311 1e−100 90 51 1 700201311H1SATMON003 g1778820 BLASTN 960 1e−97 89 52 1 700265914H1 SATMON017g1778820 BLASTN 1275 1e−97 91 53 1 700205948H1 SATMON003 g1778820 BLASTN983 1e−96 93 54 1 LIB189-003-Q1-E1-A6 LIB189 g450548 BLASTN 1262 1e−9686 55 1 LIB143-017-Q1-E1-B9 LIB143 g1778820 BLASTN 779 1e−95 86 56 1700097434H1 SATMON009 g450548 BLASTN 1247 1e−95 90 57 1 700089089H1SATMON011 g450548 BLASTN 1251 1e−95 89 58 1 700071949H1 SATMON007g1778820 BLASTN 1256 1e−95 87 59 1 700047892H1 SATMON003 g1778820 BLASTN1236 1e−94 91 60 1 700077246H1 SATMON007 g450548 BLASTN 1237 1e−94 89 611 700085681H1 SATMON011 g1778820 BLASTN 1237 1e−94 89 62 1LIB3067-010-Q1-K1-A7 LIB3067 g1778820 BLASTN 1244 1e−94 83 63 1700619961H1 SATMON034 g450548 BLASTN 962 1e−93 89 64 1 700087395H1SATMON011 g450548 BLASTN 1040 1e−93 89 65 1 700240431H1 SATMON010g1778820 BLASTN 1156 1e−93 92 66 1 700053822H1 SATMON011 g1778820 BLASTN1224 1e−93 91 67 1 700103856H1 SATMON010 g450548 BLASTN 1224 1e−93 88 681 700083703H1 SATMON011 g450548 BLASTN 1229 1e−93 89 69 1 700087360H1SATMON011 g450548 BLASTN 1234 1e−93 90 70 1 700104713H1 SATMON010g450548 BLASTN 1188 1e−92 87 71 1 700265996H1 SATMON017 g450548 BLASTN1214 1e−92 88 72 1 700264870H1 SATMON017 g450548 BLASTN 945 1e−91 89 731 700219348H1 SATMON011 g1778820 BLASTN 1203 1e−91 91 74 1 700095527H1SATMON008 g450548 BLASTN 1153 1e−90 88 75 1 LIB3068-023-Q1-K1-G5 LIB3068g1778820 BLASTN 1187 1e−90 89 76 1 700102547H1 SATMON010 g1778820 BLASTN1194 1e−90 88 77 1 700332179H1 SATMON019 g450548 BLASTN 1054 1e−89 88 781 700405470H1 SATMON029 g450548 BLASTN 1071 1e−89 90 79 1LIB3069-043-Q1-K1-G2 LIB3069 g450548 BLASTN 1175 1e−89 84 80 1700451332H1 SATMON028 g450548 BLASTN 1176 1e−89 90 81 1 700028108H1SATMON003 g450548 BLASTN 1180 1e−89 89 82 1 700076154H1 SATMON007g1778820 BLASTN 1180 1e−89 89 83 1 700089770H1 SATMON011 g450548 BLASTN1185 1e−89 89 84 1 700051882H1 SATMON003 g1778820 BLASTN 844 1e−88 91 851 700242942H1 SATMON010 g1778820 BLASTN 1163 1e−88 91 86 1 700094996H1SATMON008 g1778820 BLASTN 1166 1e−88 88 87 1 700476239H1 SATMON025g450548 BLASTN 1166 1e−88 89 88 1 700219530H1 SATMON011 g1778820 BLASTN1005 1e−87 91 89 1 700049726H1 SATMON003 g1778820 BLASTN 1041 1e−87 8990 1 700096904H1 SATMON008 g1778820 BLASTN 1157 1e−87 86 91 1700344026H1 SATMON021 g450548 BLASTN 1158 1e−87 88 92 1 700105590H1SATMON010 g1778820 BLASTN 1139 1e−86 89 93 1 700465283H1 SATMON025g450548 BLASTN 1145 1e−86 89 94 1 700104341H1 SATMON010 g450548 BLASTN1146 1e−86 87 95 1 LIB3062-021-Q1-K1-F12 LIB3062 g1778820 BLASTN 9231e−85 78 96 1 700105246H1 SATMON010 g450548 BLASTN 930 1e−85 89 97 1700072146H1 SATMON007 g1778820 BLASTN 1131 1e−85 91 98 1 700028553H1SATMON003 g1778820 BLASTN 1132 1e−85 88 99 1 700451419H1 SATMON028g450548 BLASTN 1133 1e−85 89 100 1 700103241H1 SATMON010 g450548 BLASTN1134 1e−85 90 101 1 700612549H1 SATMON033 g450548 BLASTN 1134 1e−85 89102 1 700050434H1 SATMON003 g450548 BLASTN 723 1e−84 89 103 1700094764H1 SATMON008 g1778820 BLASTN 977 1e−84 86 104 1 700163030H1SATMON013 g1778820 BLASTN 1011 1e−84 93 105 1 700466542H1 SATMON025g450548 BLASTN 1085 1e−84 89 106 1 700047380H1 SATMON003 g450548 BLASTN1096 1e−84 89 107 1 700456429H1 SATMON029 g450548 BLASTN 1118 1e−84 88108 1 700096049H1 SATMON008 g1778820 BLASTN 1122 1e−84 86 109 1700075906H1 SATMON007 g1778820 BLASTN 838 1e−83 86 110 1LIB3067-048-Q1-K1-G3 LIB3067 g1778820 BLASTN 1045 1e−83 86 111 1700214043H1 SATMON016 g450548 BLASTN 1103 1e−83 88 112 1 700096909H1SATMON008 g1778820 BLASTN 1103 1e−83 90 113 1 701184635H1 SATMONN06g1778820 BLASTN 1104 1e−83 87 114 1 700158969H1 SATMON012 g1778820BLASTN 1107 1e−83 93 115 1 700475902H1 SATMON025 g450548 BLASTN 11121e−83 89 116 1 700207076H1 SATMON003 g1778820 BLASTN 1113 1e−83 90 117 1700161901H1 SATMON012 g1778820 BLASTN 1091 1e−82 91 118 1 700452183H1SATMON028 g450548 BLASTN 1095 1e−82 88 119 1 700452326H1 SATMON028g450548 BLASTN 1098 1e−82 88 120 1 700106918H1 SATMON010 g450548 BLASTN1101 1e−82 88 121 1 700220146H1 SATMON011 g450548 BLASTN 1083 1e−81 89122 1 700221060H1 SATMON011 g450548 BLASTN 1084 1e−81 88 123 1700243344H1 SATMON010 g1778820 BLASTN 1084 1e−81 92 124 1 700475790H1SATMON025 g450548 BLASTN 1084 1e−81 89 125 1 700452691H1 SATMON028g1778820 BLASTN 1086 1e−81 87 126 1 700243733H1 SATMON010 g450548 BLASTN1086 1e−81 90 127 1 LIB3059-018-Q1-K1-D10 LIB3059 g1778820 BLASTN 8301e−80 91 128 1 700165958H1 SATMON013 g450548 BLASTN 1070 1e−80 89 129 1700082659H1 SATMON011 g1778820 BLASTN 1071 1e−80 88 130 1 700172557H1SATMON013 g1778820 BLASTN 1074 1e−80 92 131 1 700221526H1 SATMON011g450548 BLASTN 1075 1e−80 88 132 1 700456696H1 SATMON029 g450548 BLASTN1077 1e−80 89 133 1 700430776H1 SATMONN01 g1778820 BLASTN 574 1e−79 90134 1 LIB3062-025-Q1-K1-B11 LIB3062 g450548 BLASTN 585 1e−79 85 135 1700077126H1 SATMON007 g1778820 BLASTN 743 1e−79 88 136 1 700160310H1SATMON012 g1778820 BLASTN 752 1e−79 91 137 1 700030466H1 SATMON003g1778820 BLASTN 757 1e−79 83 138 1 700210204H1 SATMON016 g1778820 BLASTN844 1e−79 91 139 1 700094716H1 SATMON008 g1778820 BLASTN 857 1e−79 86140 1 700157051H1 SATMON012 g1778820 BLASTN 937 1e−79 89 141 1700241415H1 SATMON010 g450548 BLASTN 1055 1e−79 90 142 1 700160127H1SATMON012 g450548 BLASTN 1056 1e−79 88 143 1 700205485H1 SATMON003g1778820 BLASTN 1059 1e−79 90 144 1 700475930H1 SATMON025 g450548 BLASTN1060 1e−79 89 145 1 700161902H1 SATMON012 g1778820 BLASTN 1060 1e−79 91146 1 700213573H1 SATMON016 g450548 BLASTN 1061 1e−79 89 147 1700207677H1 SATMON016 g450548 BLASTN 1061 1e−79 88 148 1 700581117H1SATMON031 g450548 BLASTN 1065 1e−79 88 149 1 700451418H1 SATMON028g450548 BLASTN 546 1e−78 89 150 1 700212606H1 SATMON016 g1778820 BLASTN1044 1e−78 85 151 1 700222605H1 SATMON011 g1778820 BLASTN 1047 1e−78 86152 1 700050542H1 SATMON003 g1778820 BLASTN 1053 1e−78 86 153 1700450709H1 SATMON028 g450548 BLASTN 834 1e−77 88 154 1 700213368H1SATMON016 g450548 BLASTN 861 1e−77 89 155 1 700095926H1 SATMON008g1778820 BLASTN 1030 1e−77 86 156 1 700261686H1 SATMON017 g1778820BLASTN 1034 1e−77 89 157 1 700235993H1 SATMON010 g450548 BLASTN 10371e−77 88 158 1 700466152H1 SATMON025 g450548 BLASTN 1038 1e−77 89 159 1700266410H1 SATMON017 g1778820 BLASTN 1041 1e−77 85 160 1 700216991H1SATMON016 g1778820 BLASTN 491 1e−76 92 161 1 700464957H1 SATMON025g450548 BLASTN 607 1e−76 88 162 1 700160463H1 SATMON012 g1778820 BLASTN1018 1e−76 91 163 1 700158224H1 SATMON012 g1778820 BLASTN 1018 1e−76 92164 1 700469971H1 SATMON025 g1778820 BLASTN 1020 1e−76 86 165 1700222731H1 SATMON011 g450548 BLASTN 1022 1e−76 88 166 1 700087434H1SATMON011 g1778820 BLASTN 1023 1e−76 86 167 1 700094482H1 SATMON008g1778820 BLASTN 1023 1e−76 86 168 1 700235976H1 SATMON010 g450548 BLASTN1028 1e−76 89 169 1 700088266H1 SATMON011 g1778820 BLASTN 983 1e−75 92170 1 700075882H1 SATMON007 g1778820 BLASTN 1009 1e−75 83 171 1700243324H1 SATMON010 g1778820 BLASTN 1012 1e−75 86 172 1 700159625H2SATMON012 g450548 BLASTN 1015 1e−75 90 173 1 700204422H1 SATMON003g1778820 BLASTN 1015 1e−75 88 174 1 700048228H1 SATMON003 g1778820BLASTN 741 1e−74 85 175 1 700072474H1 SATMON007 g1778820 BLASTN 7991e−74 85 176 1 700477790H1 SATMON025 g450548 BLASTN 960 1e−74 89 177 1LIB3059-022-Q1-K1-A9 LIB3059 g1778820 BLASTN 995 1e−74 86 178 1700241633H1 SATMON010 g450548 BLASTN 997 1e−74 89 179 1 700093978H1SATMON008 g1778820 BLASTN 999 1e−74 89 180 1 700238569H1 SATMON010g450548 BLASTN 844 1e−73 88 181 1 700455160H1 SATMON029 g450548 BLASTN849 1e−73 86 182 1 700262745H1 SATMON017 g1778820 BLASTN 984 1e−73 85183 1 700405029H1 SATMON027 g1778820 BLASTN 990 1e−73 90 184 1700195230H1 SATMON014 g450548 BLASTN 991 1e−73 88 185 1 700264753H1SATMON017 g1778820 BLASTN 993 1e−73 90 186 1 700050142H1 SATMON003g1778820 BLASTN 433 1e−72 86 187 1 700452677H1 SATMON028 g450548 BLASTN919 1e−72 87 188 1 700168870H1 SATMON013 g1778820 BLASTN 971 1e−72 90189 1 700023193H1 SATMON003 g450548 BLASTN 972 1e−72 88 190 1700086582H1 SATMON011 g1778820 BLASTN 980 1e−72 84 191 1 700457710H1SATMON029 g450548 BLASTN 982 1e−72 87 192 1 700477601H1 SATMON025g450548 BLASTN 635 1e−71 88 193 1 700377730H1 SATMON019 g1778820 BLASTN691 1e−71 87 194 1 700169782H1 SATMON013 g1778820 BLASTN 960 1e−71 91195 1 700461113H1 SATMON033 g1778820 BLASTN 962 1e−71 85 196 1700258669H1 SATMON017 g1778820 BLASTN 963 1e−71 90 197 1 700454253H1SATMON029 g1778820 BLASTN 964 1e−71 85 198 1 700092376H1 SATMON008g1778820 BLASTN 780 1e−70 83 199 1 700242885H1 SATMON010 g960356 BLASTN946 1e−70 86 200 1 700094333H1 SATMON008 g1778820 BLASTN 947 1e−70 86201 1 700575124H1 SATMON030 g450548 BLASTN 948 1e−70 82 202 1700072544H1 SATMON007 g1778820 BLASTN 949 1e−70 86 203 1LIB143-046-Q1-E1-B5 LIB143 g450548 BLASTN 951 1e−70 87 204 1 700096534H1SATMON008 g1778820 BLASTN 956 1e−70 85 205 1 700262906H1 SATMON017g1778820 BLASTN 937 1e−69 87 206 1 700094478H1 SATMON008 g1778820 BLASTN939 1e−69 90 207 1 700172985H1 SATMON013 g450548 BLASTN 939 1e−69 88 2081 700097938H1 SATMON009 g1778820 BLASTN 941 1e−69 85 209 1LIB3069-006-Q1-K1-D5 LIB3069 g450548 BLASTN 943 1e−69 88 210 1700093125H1 SATMON008 g1778820 BLASTN 945 1e−69 88 211 1 700424156H1SATMONN01 g450548 BLASTN 752 1e−68 85 212 1 700405486H1 SATMON029g450548 BLASTN 864 1e−68 91 213 1 LIB189-026-Q1-E1-H12 LIB189 g1778820BLASTN 879 1e−68 87 214 1 700106172H1 SATMON010 g1778820 BLASTN 9251e−68 85 215 1 700096890H1 SATMON008 g1778820 BLASTN 926 1e−68 90 216 1700154404H1 SATMON007 g450548 BLASTN 928 1e−68 89 217 1 700468337H1SATMON025 g450548 BLASTN 445 1e−67 88 218 1 700158827H1 SATMON012g1778820 BLASTN 522 1e−67 87 219 1 700241744H1 SATMON010 g1778820 BLASTN575 1e−67 85 220 1 700624633H1 SATMON034 g960356 BLASTN 644 1e−67 85 2211 700154564H1 SATMON007 g1778820 BLASTN 735 1e−67 85 222 1 700172424H1SATMON013 g450548 BLASTN 912 1e−67 90 223 1 700096287H1 SATMON008g1778820 BLASTN 914 1e−67 86 224 1 700207638H1 SATMON016 g1778820 BLASTN914 1e−67 86 225 1 700093735H1 SATMON008 g960356 BLASTN 920 1e−67 89 2261 700159494H1 SATMON012 g1778820 BLASTN 899 1e−66 89 227 1 700236013H1SATMON010 g1778820 BLASTN 900 1e−66 83 228 1 700220267H1 SATMON011g1778820 BLASTN 439 1e−65 84 229 1 700477578H1 SATMON025 g450548 BLASTN576 1e−65 90 230 1 700047743H1 SATMON003 g1778820 BLASTN 601 1e−65 82231 1 700343041H1 SATMON021 g1778820 BLASTN 633 1e−65 86 232 1700159886H1 SATMON012 g450548 BLASTN 803 1e−65 85 233 1 700570242H1SATMON030 g450548 BLASTN 834 1e−65 85 234 1 700021930H1 SATMON001g1778820 BLASTN 888 1e−65 87 235 1 700454959H1 SATMON029 g450548 BLASTN890 1e−65 89 236 1 700171336H1 SATMON013 g1778820 BLASTN 890 1e−65 89237 1 700105361H1 SATMON010 g1778820 BLASTN 806 1e−64 87 238 1LIB3068-031-Q1-K1-B1 LIB3068 g450548 BLASTN 853 1e−64 89 239 1700236324H1 SATMON010 g450548 BLASTN 875 1e−64 89 240 1 700150620H1SATMON007 g450548 BLASTN 880 1e−64 87 241 1 700220648H1 SATMON011g450548 BLASTN 881 1e−64 90 242 1 700150733H1 SATMON007 g450548 BLASTN881 1e−64 85 243 1 700157367H1 SATMON012 g1778820 BLASTN 882 1e−64 85244 1 700259676H1 SATMON017 g1778820 BLASTN 885 1e−64 88 245 1700616490H1 SATMON033 g450548 BLASTN 886 1e−64 82 246 1 700102511H1SATMON010 g450548 BLASTN 695 1e−63 89 247 1 700202805H1 SATMON003g1778820 BLASTN 817 1e−63 92 248 1 700105685H1 SATMON010 g1778820 BLASTN866 1e−63 84 249 1 700106113H1 SATMON010 g450548 BLASTN 873 1e−63 91 2501 700444778H1 SATMON027 g1778820 BLASTN 343 1e−62 84 251 1 700103584H1SATMON010 g450548 BLASTN 521 1e−62 86 252 1 LIB189-008-Q1-E1-D9 LIB189g1778820 BLASTN 850 1e−62 91 253 1 700155684H1 SATMON007 g1778820 BLASTN852 1e−62 85 254 1 700261639H1 SATMON017 g1778820 BLASTN 853 1e−62 89255 1 700158367H1 SATMON012 g450548 BLASTN 856 1e−62 81 256 1700153242H1 SATMON007 g1778820 BLASTN 859 1e−62 90 257 1 700210738H1SATMON016 g1778820 BLASTN 859 1e−62 90 258 1 700203008H1 SATMON003g450548 BLASTN 698 1e−61 86 259 1 700206081H1 SATMON003 g1778820 BLASTN840 1e−61 92 260 1 700028643H1 SATMON003 g1778820 BLASTN 846 1e−61 85261 1 700223914H1 SATMON011 g1778820 BLASTN 849 1e−61 84 262 1700571455H1 SATMON030 g1778820 BLASTN 378 1e−60 88 263 1 700075374H1SATMON007 g1778820 BLASTN 830 1e−60 85 264 1 700150452H1 SATMON007g450548 BLASTN 831 1e−60 89 265 1 700261318H1 SATMON017 g1778820 BLASTN831 1e−60 83 266 1 700616390H1 SATMON033 g450548 BLASTN 835 1e−60 92 2671 700208718H1 SATMON016 g450548 BLASTN 561 1e−59 89 268 1 700448948H1SATMON028 g1778820 BLASTN 653 1e−59 83 269 1 LIB3060-035-Q1-K1-H3LIB3060 g450548 BLASTN 734 1e−59 85 270 1 700049753H1 SATMON003 g450548BLASTN 769 1e−59 90 271 1 700154489H1 SATMON007 g1778820 BLASTN 8141e−59 83 272 1 700170783H1 SATMON013 g1778820 BLASTN 816 1e−59 87 273 1700237571H1 SATMON010 g450548 BLASTN 817 1e−59 89 274 1 700154872H1SATMON007 g1778820 BLASTN 822 1e−59 85 275 1 700025620H1 SATMON004g1778820 BLASTN 686 1e−58 84 276 1 700158255H1 SATMON012 g450548 BLASTN803 1e−58 86 277 1 700159282H1 SATMON012 g450548 BLASTN 805 1e−58 83 2781 700222171H1 SATMON011 g1778820 BLASTN 807 1e−58 89 279 1 700205003H1SATMON003 g1778820 BLASTN 441 1e−57 86 280 1 700049209H1 SATMON003g1778820 BLASTN 443 1e−57 90 281 1 700016430H1 SATMON001 g1778820 BLASTN492 1e−57 86 282 1 700212936H1 SATMON016 g450548 BLASTN 564 1e−57 86 2831 700156939H1 SATMON012 g1778820 BLASTN 801 1e−57 84 284 1 700222183H1SATMON011 g1778820 BLASTN 561 1e−56 83 285 1 700203453H1 SATMON003g1778820 BLASTN 784 1e−56 90 286 1 700167708H1 SATMON013 g1778820 BLASTN560 1e−55 83 287 1 700214356H1 SATMON016 g1778820 BLASTN 693 1e−55 87288 1 700475377H1 SATMON025 g450548 BLASTN 705 1e−55 90 289 1700029625H1 SATMON003 g450548 BLASTN 712 1e−55 88 290 1 700202091H1SATMON003 g1778820 BLASTN 774 1e−55 90 291 1 700264594H1 SATMON017g450548 BLASTN 775 1e−55 89 292 1 700074969H1 SATMON007 g450548 BLASTN777 1e−55 87 293 1 LIB3068-005-Q1-K1-A9 LIB3068 g1778820 BLASTN 3891e−54 80 294 1 700207147H1 SATMON017 g1778820 BLASTN 537 1e−54 88 295 1LIB189-004-Q1-E1-F11 LIB189 g960356 BLASTN 757 1e−54 87 296 1700171222H1 SATMON013 g1778820 BLASTN 757 1e−54 93 297 1 700239903H1SATMON010 g1778820 BLASTN 760 1e−54 84 298 1 700048208H1 SATMON003g1778820 BLASTN 762 1e−54 90 299 1 700264085H1 SATMON017 g450548 BLASTN764 1e−54 87 300 1 700155761H1 SATMON007 g1778820 BLASTN 579 1e−53 85301 1 700216516H1 SATMON016 g450548 BLASTN 380 1e−52 89 302 1LIB3068-044-Q1-K1-F12 LIB3068 g450548 BLASTN 492 1e−52 73 303 1700344420H1 SATMON021 g450548 BLASTN 534 1e−52 79 304 1 700219767H1SATMON011 g450548 BLASTN 732 1e−52 88 305 1 700153757H1 SATMON007g1778820 BLASTN 737 1e−52 89 306 1 700165841H1 SATMON013 g450548 BLASTN738 1e−52 90 307 1 700381966H1 SATMON023 g1778820 BLASTN 740 1e−52 86308 1 700170427H1 SATMON013 g450548 BLASTN 687 1e−51 89 309 1700094344H1 SATMON008 g1778820 BLASTN 725 1e−51 93 310 1 700052248H1SATMON003 g1778820 BLASTN 726 1e−51 84 311 1 700050628H1 SATMON003g450548 BLASTN 726 1e−51 89 312 1 700223031H1 SATMON011 g1778820 BLASTN729 1e−51 89 313 1 700475686H1 SATMON025 g450548 BLASTN 660 1e−50 89 3141 700170325H1 SATMON013 g2305013 BLASTN 709 1e−50 82 315 1 700221107H1SATMON011 g450548 BLASTN 698 1e−49 89 316 1 700612589H1 SATMON033g960356 BLASTN 703 1e−49 89 317 1 700153770H1 SATMON007 g1778820 BLASTN705 1e−49 88 318 1 LIB3078-006-Q1-K1-E7 LIB3078 g1778820 BLASTN 7051e−49 79 319 1 700239724H1 SATMON010 g1778820 BLASTN 507 1e−48 78 320 1700356777H1 SATMON024 g1778820 BLASTN 683 1e−48 86 321 1 700241568H1SATMON010 g960356 BLASTN 691 1e−48 88 322 1 700051294H1 SATMON003g1778820 BLASTN 462 1e−47 88 323 1 700343661H1 SATMON021 g450548 BLASTN507 1e−47 81 324 1 700029419H1 SATMON003 g1778820 BLASTN 547 1e−47 87325 1 700165936H1 SATMON013 g1778820 BLASTN 671 1e−47 83 326 1700026152H1 SATMON003 g960356 BLASTN 673 1e−47 89 327 1 700075671H1SATMON007 g450548 BLASTN 429 1e−46 89 328 1 700156674H1 SATMON012g1778820 BLASTN 659 1e−46 91 329 1 700094981H1 SATMON008 g1778820 BLASTN662 1e−46 85 330 1 700092879H1 SATMON008 g1778820 BLASTN 668 1e−46 87331 1 700240685H1 SATMON010 g1778820 BLASTN 484 1e−45 84 332 1700150286H1 SATMON007 g1778820 BLASTN 647 1e−45 82 333 1 700104990H1SATMON010 g450548 BLASTN 652 1e−45 89 334 1 700203829H1 SATMON003g450548 BLASTN 652 1e−45 87 335 1 700153718H1 SATMON007 g167961 BLASTN656 1e−45 91 336 1 700050841H1 SATMON003 g450548 BLASTN 571 1e−44 86 3371 700268037H1 SATMON017 g2305013 BLASTN 636 1e−44 86 338 1 700153630H1SATMON007 g1778820 BLASTN 637 1e−44 82 339 1 700475317H1 SATMON025g450548 BLASTN 530 1e−43 87 340 1 701163127H1 SATMONN04 g960356 BLASTN626 1e−43 88 341 1 700203688H1 SATMON003 g960356 BLASTN 627 1e−43 89 3421 700049893H1 SATMON003 g1778820 BLASTN 506 1e−42 90 343 1 700449155H1SATMON028 g1778820 BLASTN 443 1e−41 85 344 1 701183780H1 SATMONN06g450548 BLASTN 558 1e−41 86 345 1 700242162H1 SATMON010 g1778820 BLASTN600 1e−41 90 346 1 700466994H1 SATMON025 g450548 BLASTN 604 1e−41 91 3471 700259823H1 SATMON017 g450548 BLASTN 607 1e−41 83 348 1 700346242H1SATMON021 g450548 BLASTN 608 1e−41 86 349 1 700156395H1 SATMON007g1778820 BLASTN 609 1e−41 90 350 1 700236835H1 SATMON010 g1778820 BLASTN335 1e−40 81 351 1 700172385H1 SATMON013 g1778820 BLASTN 397 1e−40 87352 1 700210466H1 SATMON016 g1778820 BLASTN 586 1e−40 81 353 1700257303H1 SATMON017 g1778820 BLASTN 589 1e−40 81 354 1LIB3067-027-Q1-K1-G1 LIB3067 g1778820 BLASTN 623 1e−40 87 355 1LIB3066-025-Q1-K1-D1 LIB3066 g450548 BLASTN 524 1e−39 85 356 1700106853H1 SATMON010 g450548 BLASTN 580 1e−39 86 357 1 700160540H1SATMON012 g960356 BLASTN 581 1e−39 88 358 1 700157780H1 SATMON012g960356 BLASTN 581 1e−39 88 359 1 700149801H1 SATMON007 g450548 BLASTN565 1e−38 86 360 1 700353243H1 SATMON024 g450548 BLASTN 570 1e−38 86 3611 700166171H1 SATMON013 g1778820 BLASTN 254 1e−37 79 362 1 700142509H1SATMON012 g960356 BLASTN 556 1e−37 88 363 1 700242131H1 SATMON010g450548 BLASTN 560 1e−37 85 364 1 700455643H1 SATMON029 g450548 BLASTN539 1e−36 86 365 1 700150248H1 SATMON007 g1778820 BLASTN 547 1e−36 87366 1 700208549H1 SATMON016 g450548 BLASTN 559 1e−36 88 367 1700027193H1 SATMON003 g450548 BLASTN 529 1e−35 86 368 1 700221390H1SATMON011 g450548 BLASTN 530 1e−35 85 369 1 700455647H1 SATMON029g450548 BLASTN 531 1e−35 85 370 1 700260103H1 SATMON017 g1778820 BLASTN531 1e−35 80 371 1 700167344H1 SATMON013 g1778820 BLASTN 536 1e−35 88372 1 700089913H1 SATMON011 g960356 BLASTN 546 1e−35 88 373 1700570573H1 SATMON030 g450548 BLASTN 300 1e−34 89 374 1 700169889H1SATMON013 g1778820 BLASTN 519 1e−34 89 375 1 700262857H1 SATMON017g1778820 BLASTN 521 1e−34 87 376 1 700142644H2 SATMON013 g1778820 BLASTN522 1e−34 87 377 1 700224417H1 SATMON011 g450548 BLASTN 522 1e−34 91 3781 700073882H1 SATMON007 g450548 BLASTN 531 1e−34 87 379 1 700085803H1SATMON011 g450548 BLASTN 338 1e−33 89 380 1 700162323H1 SATMON012g1778820 BLASTN 513 1e−33 86 381 1 700468306H1 SATMON025 g450548 BLASTN519 1e−33 89 382 1 700443224H2 SATMON026 g450548 BLASTN 275 1e−32 83 3831 LIB3066-054-Q1-K1-E3 LIB3066 g450548 BLASTN 495 1e−32 87 384 1700211827H1 SATMON016 g1778820 BLASTN 489 1e−31 80 385 1 700048741H1SATMON003 g450548 BLASTN 500 1e−31 88 386 1 700613620H1 SATMON033g1778820 BLASTN 385 1e−30 88 387 1 700029203H1 SATMON003 g1778820 BLASTN461 1e−29 84 388 1 700378431H1 SATMON020 g450548 BLASTN 466 1e−29 89 3891 700455641H1 SATMON029 g450548 BLASTN 472 1e−29 85 390 1 700447867H1SATMON027 g960356 BLASTN 473 1e−29 88 391 1 700447511H1 SATMON027g450548 BLASTN 474 1e−29 88 392 1 700025851H1 SATMON003 g450548 BLASTN479 1e−29 88 393 1 LIB3066-024-Q1-K1-H4 LIB3066 g1778820 BLASTN 4811e−29 81 394 1 LIB143-025-Q1-E1-C4 LIB143 g450549 BLASTX 64 1e−28 70 3951 700242282H1 SATMON010 g1778820 BLASTN 280 1e−28 85 396 1 700087244H1SATMON011 g450548 BLASTN 464 1e−28 88 397 1 700458127H1 SATMON029g450548 BLASTN 238 1e−27 82 398 1 700025767H1 SATMON003 g1778820 BLASTN438 1e−26 86 399 1 700235401H1 SATMON010 g450548 BLASTN 442 1e−26 88 4001 700029457H1 SATMON003 g450548 BLASTN 446 1e−26 89 401 1 700266174H1SATMON017 g450548 BLASTN 447 1e−26 89 402 1 700349254H1 SATMON023g1778820 BLASTN 315 1e−25 85 403 1 LIB3068-041-Q1-K1-H6 LIB3068 g450548BLASTN 438 1e−25 86 404 1 700092889H1 SATMON008 g1778820 BLASTN 3971e−24 92 405 1 700049044H1 SATMON003 g1778820 BLASTN 425 1e−24 90 406 1700202710H1 SATMON003 g960357 BLASTX 114 1e−19 97 407 1 700155117H1SATMON007 g450548 BLASTN 314 1e−19 90 408 1 700449619H1 SATMON028g1778820 BLASTN 341 1e−19 89 409 1 700150336H1 SATMON007 g1778820 BLASTN343 1e−19 87 410 1 700166223H1 SATMON013 g960357 BLASTX 181 1e−18 97 4111 700153796H1 SATMON007 g2305013 BLASTN 300 1e−16 82 412 1 700053266H1SATMON008 g450548 BLASTN 292 1e−15 83 413 1 700405227H1 SATMON028g450548 BLASTN 313 1e−15 90 414 1 700397410H1 SATMONN01 g17262 BLASTX147 1e−14 96 415 1 700211524H1 SATMON016 g1033190 BLASTX 153 1e−14 100416 1 700281415H2 SATMON019 g2315140 BLASTX 139 1e−13 89 417 1700429873H1 SATMONN01 g16961 BLASTX 133 1e−12 94 418 1 700239660H1SATMON010 g450548 BLASTN 245 1e−12 86 419 1 700213596H1 SATMON016g450548 BLASTN 258 1e−12 95 420 1 700152367H1 SATMON007 g450548 BLASTN273 1e−12 88 421 1 700452014H1 SATMON028 g17262 BLASTX 76 1e−11 72 422 1700357106H1 SATMON024 g1724104 BLASTX 132 1e−11 100 423 1 700468611H1SATMON025 g450549 BLASTX 93 1e−10 93 424 1 700213526H1 SATMON016 g450549BLASTX 127 1e−10 96 425 1 700267065H1 SATMON017 g450549 BLASTX 130 1e−1096 426 1 700152044H1 SATMON007 g450549 BLASTX 88 1e−8 93 427 1700159090H1 SATMON012 g169665 BLASTX 113 1e−8 81 428 1 700266734H1SATMON017 g450549 BLASTX 119 1e−8 96 429 1 700405367H1 SATMON029g1778820 BLASTN 231 1e−8 65 1635 -700555532 700555532H1 SOYMON001g429103 BLASTN 920 1e−67 84 1636 -700649594 700649594H1 SOYMON003g609559 BLASTX 186 1e−23 85 1637 -700750590 700750590H1 SOYMON014g609224 BLASTN 363 1e−40 79 1638 -700755802 700755802H1 SOYMON014g609224 BLASTN 479 1e−43 74 1639 -700869211 700869211H1 SOYMON016g169665 BLASTX 146 1e−13 92 1640 -700891960 700891960H1 SOYMON024g726031 BLASTN 589 1e−56 82 1641 -700900377 700900377H1 SOYMON027g1655577 BLASTN 235 1e−8 78 1642 -700902427 700902427H1 SOYMON027g1655576 BLASTX 151 1e−13 76 1643 -700941686 700941686H1 SOYMON024g497899 BLASTN 442 1e−26 89 1644 -700952418 700952418H1 SOYMON022g726030 BLASTX 146 1e−17 83 1645 -700979651 700979651H2 SOYMON009g1655579 BLASTN 1006 1e−75 84 1646 -700982809 700982809H1 SOYMON009g1655579 BLASTN 945 1e−69 82 1647 -700982867 700982867H1 SOYMON009g1127582 BLASTN 868 1e−63 78 1648 -701056884 701056884H1 SOYMON032g609556 BLASTN 726 1e−51 84 1649 -701117318 701117318H1 SOYMON037g609224 BLASTN 451 1e−37 80 1650 -701118224 701118224H1 SOYMON037g2305013 BLASTN 285 1e−19 72 1651 -701121264 701121264H1 SOYMON037g166873 BLASTN 238 1e−8 82 1652 -701122908 701122908H1 SOYMON037 g16508BLASTN 444 1e−26 83 1653 -701128589 701128589H1 SOYMON037 g16508 BLASTN539 1e−36 90 1654 -GM12798 LIB3049-039-Q1-E1-F2 LIB3049 g16508 BLASTN578 1e−37 73 1655 -GM14331 LIB3049-055-Q1-E1-F5 LIB3049 g167961 BLASTN497 1e−30 62 1656 -GM30881 LIB3050-005-Q1-K1-G1 LIB3050 g1655577 BLASTN543 1e−34 82 1657 -GM30911 LIB3050-005-Q1-K1-B8 LIB3050 g1655577 BLASTN387 1e−26 71 1658 -GM33921 LIB3051-028-Q1-K1-A9 LIB3051 g16508 BLASTN338 1e−32 73 1659 12644 701131794H1 SOYMON038 g429107 BLASTN 877 1e−6484 1660 12644 701142515H1 SOYMON038 g429107 BLASTN 871 1e−63 84 166112644 700888494H1 SOYMON024 g429107 BLASTN 662 1e−46 83 1662 16LIB3030-009-Q1-B1-C1 LIB3030 g429105 BLASTN 1439 1e−119 84 1663 16LIB3051-106-Q1-K1-B5 LIB3051 g609224 BLASTN 1516 1e−117 86 1664 16LIB3050-023-Q1-K1-A12 LIB3050 g1724103 BLASTN 1388 1e−106 83 1665 16LIB3028-003-Q1-B1-G11 LIB3028 g609224 BLASTN 1313 1e−100 87 1666 16LIB3054-010-Q1-N1-E2 LIB3054 g609224 BLASTN 920 1e−95 87 1667 16700651294H1 SOYMON003 g609224 BLASTN 672 1e−94 86 1668 16LIB3027-007-Q1-B1-G3 LIB3027 g16508 BLASTN 891 1e−93 83 1669 16LIB3053-013-Q1-N1-H9 LIB3053 g609224 BLASTN 996 1e−93 87 1670 16LIB3051-061-Q1-K1-C7 LIB3051 g16508 BLASTN 1195 1e−93 86 1671 16LIB3065-005-Q1-N1-A6 LIB3065 g609224 BLASTN 707 1e−91 78 1672 16LIB3050-008-Q1-E1-E3 LIB3050 g609224 BLASTN 1102 1e−87 87 1673 16LIB3051-011-Q1-E1-B2 LIB3051 g16508 BLASTN 1153 1e−87 82 1674 16LIB3030-010-Q1-B1-F8 LIB3030 g609224 BLASTN 988 1e−86 83 1675 16700652104H1 SOYMON003 g1655577 BLASTN 990 1e−86 83 1676 16 701205279H1SOYMON035 g609556 BLASTN 1125 1e−85 86 1677 16 700662183H1 SOYMON005g609224 BLASTN 678 1e−83 84 1678 16 LIB3051-039-Q1-K1-F5 LIB3051 g169664BLASTN 1104 1e−83 86 1679 16 700557616H1 SOYMON001 g609556 BLASTN 11111e−83 86 1680 16 LIB3030-002-Q1-B1-C6 LIB3030 g16508 BLASTN 1113 1e−8383 1681 16 700865235H1 SOYMON016 g1724103 BLASTN 1090 1e−82 84 1682 16700563340H1 SOYMON002 g609224 BLASTN 911 1e−80 86 1683 16LIB3040-044-Q1-E1-D7 LIB3040 g166873 BLASTN 1052 1e−80 82 1684 16LIB3040-060-Q1-E1-D9 LIB3040 g609224 BLASTN 1071 1e−80 84 1685 16LIB3049-001-Q1-E1-F12 LIB3049 g16508 BLASTN 1074 1e−80 84 1686 16LIB3028-006-Q1-B1-F1 LIB3028 g16508 BLASTN 857 1e−79 83 1687 16LIB3049-033-Q1-E1-G5 LIB3049 g2315139 BLASTN 933 1e−79 81 1688 16700978240H1 SOYMON009 g609224 BLASTN 984 1e−79 88 1689 16 700980802H1SOYMON009 g862999 BLASTN 1060 1e−79 85 1690 16 700755908H1 SOYMON014g1724103 BLASTN 1062 1e−79 88 1691 16 700562226H1 SOYMON002 g1724103BLASTN 1065 1e−79 84 1692 16 701119101H1 SOYMON037 g609224 BLASTN 10481e−78 86 1693 16 700646291H1 SOYMON012 g1655577 BLASTN 1049 1e−78 861694 16 LIB3050-022-Q1-K1-B9 LIB3050 g16508 BLASTN 1049 1e−78 83 1695 16LIB3028-030-Q1-B1-F8 LIB3028 g16508 BLASTN 1053 1e−78 83 1696 16701143128H1 SOYMON038 g609556 BLASTN 737 1e−77 86 1697 16LIB3040-041-Q1-E1-D10 LIB3040 g166873 BLASTN 861 1e−77 81 1698 16700756363H1 SOYMON014 g609556 BLASTN 1031 1e−77 88 1699 16 700729963H1SOYMON009 g1724103 BLASTN 1036 1e−77 88 1700 16 701063046H1 SOYMON033g609224 BLASTN 1037 1e−77 86 1701 16 LIB3030-005-Q1-B1-G2 LIB3030g609224 BLASTN 1038 1e−77 84 1702 16 700564331H1 SOYMON002 g609556BLASTN 1039 1e−77 84 1703 16 700562958H1 SOYMON002 g1724103 BLASTN 10401e−77 88 1704 16 LIB3039-014-Q1-E1-F7 LIB3039 g16508 BLASTN 569 1e−76 841705 16 700753632H1 SOYMON014 g497899 BLASTN 668 1e−76 83 1706 16LIB3049-048-Q1-E1-F2 LIB3049 g16508 BLASTN 722 1e−76 83 1707 16700648911H1 SOYMON003 g1655577 BLASTN 983 1e−76 83 1708 16LIB3040-027-Q1-E1-H3 LIB3040 g166873 BLASTN 1003 1e−76 81 1709 16700664905H1 SOYMON005 g609556 BLASTN 1020 1e−76 86 1710 16 701133404H1SOYMON038 g1724103 BLASTN 1023 1e−76 85 1711 16 701208780H1 SOYMON035g1655577 BLASTN 1025 1e−76 85 1712 16 700902279H1 SOYMON027 g169664BLASTN 1026 1e−76 88 1713 16 LIB3040-048-Q1-E1-G10 LIB3040 g16508 BLASTN1030 1e−76 83 1714 16 LIB3040-028-Q1-E1-A4 LIB3040 g16508 BLASTN 9791e−75 84 1715 16 700807586H1 SOYMON016 g609224 BLASTN 1006 1e−75 86 171616 700755486H1 SOYMON014 g609224 BLASTN 1006 1e−75 87 1717 16700724920H1 SOYMON009 g609224 BLASTN 1009 1e−75 87 1718 16 701007494H2SOYMON019 g1724103 BLASTN 1013 1e−75 85 1719 16 700568310H1 SOYMON002g497899 BLASTN 834 1e−74 83 1720 16 701208819H1 SOYMON035 g609224 BLASTN857 1e−74 87 1721 16 700985084H1 SOYMON009 g609224 BLASTN 872 1e−74 851722 16 701013074H1 SOYMON019 g726031 BLASTN 894 1e−74 88 1723 16700650843H1 SOYMON003 g609224 BLASTN 924 1e−74 86 1724 16 700646037H1SOYMON011 g609556 BLASTN 930 1e−74 84 1725 16 LIB3040-039-Q1-E1-H9LIB3040 g166873 BLASTN 973 1e−74 80 1726 16 700847231H1 SOYMON021g726031 BLASTN 999 1e−74 85 1727 16 700792293H1 SOYMON011 g609224 BLASTN999 1e−74 86 1728 16 701109644H1 SOYMON036 g609556 BLASTN 1004 1e−74 861729 16 701122929H1 SOYMON037 g1724103 BLASTN 459 1e−73 87 1730 16700661491H1 SOYMON005 g16508 BLASTN 540 1e−73 90 1731 16 701120070H1SOYMON037 g609224 BLASTN 786 1e−73 87 1732 16 700898895H1 SOYMON027g609556 BLASTN 820 1e−73 88 1733 16 701096959H1 SOYMON028 g429105 BLASTN839 1e−73 82 1734 16 700848356H1 SOYMON021 g497899 BLASTN 985 1e−73 851735 16 700864876H1 SOYMON016 g1724103 BLASTN 985 1e−73 88 1736 16700868456H1 SOYMON016 g609224 BLASTN 987 1e−73 86 1737 16 701210488H1SOYMON035 g1655577 BLASTN 988 1e−73 83 1738 16 700747764H1 SOYMON013g609224 BLASTN 822 1e−72 87 1739 16 701063311H1 SOYMON033 g1655577BLASTN 971 1e−72 86 1740 16 700868625H1 SOYMON016 g1724103 BLASTN 9731e−72 85 1741 16 700992344H1 SOYMON011 g726031 BLASTN 973 1e−72 85 174216 701012719H1 SOYMON019 g1724103 BLASTN 973 1e−72 85 1743 16700676769H1 SOYMON007 g609224 BLASTN 973 1e−72 87 1744 16 700946235H1SOYMON024 g609224 BLASTN 975 1e−72 85 1745 16 700891218H1 SOYMON024g1724103 BLASTN 975 1e−72 84 1746 16 700724907H1 SOYMON009 g609556BLASTN 977 1e−72 84 1747 16 700833549H1 SOYMON019 g1655577 BLASTN 9791e−72 85 1748 16 700942105H1 SOYMON024 g726031 BLASTN 980 1e−72 84 174916 700564290H1 SOYMON002 g726031 BLASTN 530 1e−71 84 1750 16 700891233H1SOYMON024 g1724103 BLASTN 746 1e−71 86 1751 16 LIB3028-025-Q1-B1-B2LIB3028 g609224 BLASTN 870 1e−71 85 1752 16 701120896H1 SOYMON037g429105 BLASTN 959 1e−71 82 1753 16 LIB3051-027-Q1-K1-A9 LIB3051 g609224BLASTN 962 1e−71 83 1754 16 701050696H1 SOYMON032 g609556 BLASTN 9651e−71 86 1755 16 700653619H1 SOYMON003 g609224 BLASTN 966 1e−71 86 175616 701047994H1 SOYMON032 g1724103 BLASTN 967 1e−71 85 1757 16701123056H1 SOYMON037 g1724103 BLASTN 968 1e−71 85 1758 16 700983479H1SOYMON009 g726031 BLASTN 653 1e−70 85 1759 16 701063733H1 SOYMON034g1655577 BLASTN 850 1e−70 85 1760 16 700738366H1 SOYMON012 g726031BLASTN 946 1e−70 85 1761 16 700866319H1 SOYMON016 g1655575 BLASTN 9471e−70 85 1762 16 700789013H2 SOYMON011 g1655577 BLASTN 948 1e−70 85 176316 700896080H1 SOYMON027 g609224 BLASTN 949 1e−70 87 1764 16LIB3039-021-Q1-E1-F12 LIB3039 g16508 BLASTN 950 1e−70 83 1765 16700898284H1 SOYMON027 g429105 BLASTN 950 1e−70 86 1766 16 700901743H1SOYMON027 g609224 BLASTN 950 1e−70 86 1767 16 700831048H1 SOYMON019g429105 BLASTN 951 1e−70 85 1768 16 701038107H1 SOYMON029 g726031 BLASTN952 1e−70 87 1769 16 700559703H1 SOYMON001 g726031 BLASTN 954 1e−70 881770 16 700848817H1 SOYMON021 g609224 BLASTN 954 1e−70 84 1771 16700896032H1 SOYMON027 g609224 BLASTN 956 1e−70 87 1772 16 700944764H1SOYMON024 g16508 BLASTN 567 1e−69 85 1773 16 701011015H1 SOYMON019g1724103 BLASTN 647 1e−69 81 1774 16 701125662H1 SOYMON037 g609224BLASTN 751 1e−69 87 1775 16 701046895H1 SOYMON032 g16508 BLASTN 7951e−69 83 1776 16 701012632H1 SOYMON019 g1655577 BLASTN 800 1e−69 85 177716 701005244H1 SOYMON019 g1724103 BLASTN 804 1e−69 82 1778 16LIB3050-023-Q1-K1-H10 LIB3050 g609224 BLASTN 899 1e−69 83 1779 16700892558H1 SOYMON024 g1655575 BLASTN 936 1e−69 85 1780 16 700988779H1SOYMON011 g16508 BLASTN 937 1e−69 82 1781 16 701203923H2 SOYMON035g429105 BLASTN 938 1e−69 86 1782 16 701010231H2 SOYMON019 g1724103BLASTN 938 1e−69 83 1783 16 701041790H1 SOYMON029 g450548 BLASTN 9391e−69 84 1784 16 700967887H1 SOYMON033 g1724103 BLASTN 940 1e−69 87 178516 701123361H1 SOYMON037 g16508 BLASTN 941 1e−69 84 1786 16 701123154H1SOYMON037 g1724103 BLASTN 942 1e−69 86 1787 16 701045767H1 SOYMON032g726031 BLASTN 942 1e−69 85 1788 16 700983288H1 SOYMON009 g609224 BLASTN945 1e−69 83 1789 16 700653053H1 SOYMON003 g609224 BLASTN 945 1e−69 861790 16 701044226H1 SOYMON032 g1655577 BLASTN 486 1e−68 83 1791 16700969927H1 SOYMON005 g497899 BLASTN 750 1e−68 85 1792 16 700547956H1SOYMON001 g609224 BLASTN 779 1e−68 87 1793 16 LIB3049-048-Q1-E1-E4LIB3049 g609224 BLASTN 827 1e−68 83 1794 16 701140780H1 SOYMON038g1655577 BLASTN 922 1e−68 85 1795 16 700849166H1 SOYMON021 g1724103BLASTN 923 1e−68 84 1796 16 700891945H1 SOYMON024 g609556 BLASTN 9251e−68 87 1797 16 700658051H1 SOYMON004 g429105 BLASTN 925 1e−68 83 179816 700942415H1 SOYMON024 g1655575 BLASTN 928 1e−68 84 1799 16701041890H1 SOYMON029 g1724103 BLASTN 930 1e−68 82 1800 16LIB3028-006-Q1-B1-H12 LIB3028 g16508 BLASTN 931 1e−68 81 1801 16700967039H1 SOYMON029 g429105 BLASTN 931 1e−68 85 1802 16 700836178H1SOYMON019 g1724103 BLASTN 933 1e−68 85 1803 16 700897558H1 SOYMON027g16508 BLASTN 737 1e−67 85 1804 16 LIB3050-004-Q1-E1-A2 LIB3050 g609224BLASTN 796 1e−67 80 1805 16 700730236H1 SOYMON009 g726031 BLASTN 8161e−67 84 1806 16 700833936H1 SOYMON019 g1724103 BLASTN 915 1e−67 84 180716 700897552H1 SOYMON027 g1655577 BLASTN 916 1e−67 84 1808 16700945440H1 SOYMON024 g609556 BLASTN 917 1e−67 84 1809 16 700961368H1SOYMON022 g169664 BLASTN 918 1e−67 88 1810 16 700789702H1 SOYMON011g609224 BLASTN 921 1e−67 86 1811 16 700786541H1 SOYMON011 g429105 BLASTN513 1e−66 84 1812 16 700987282H1 SOYMON009 g16508 BLASTN 523 1e−66 871813 16 700661002H1 SOYMON005 g2305013 BLASTN 836 1e−66 81 1814 16701148312H1 SOYMON031 g862999 BLASTN 899 1e−66 84 1815 16 700738822H1SOYMON012 g1655577 BLASTN 899 1e−66 85 1816 16 700940996H1 SOYMON024g609556 BLASTN 901 1e−66 86 1817 16 700749195H1 SOYMON013 g609556 BLASTN903 1e−66 81 1818 16 700893941H1 SOYMON024 g429105 BLASTN 903 1e−66 881819 16 700892888H1 SOYMON024 g609556 BLASTN 906 1e−66 86 1820 16700901481H1 SOYMON027 g169664 BLASTN 638 1e−65 86 1821 16 700945269H1SOYMON024 g169664 BLASTN 688 1e−65 86 1822 16 700746876H1 SOYMON013g609556 BLASTN 740 1e−65 85 1823 16 700755043H1 SOYMON014 g609556 BLASTN760 1e−65 87 1824 16 701097166H1 SOYMON028 g1655577 BLASTN 781 1e−65 821825 16 701129484H1 SOYMON037 g16508 BLASTN 898 1e−65 82 1826 16700651014H1 SOYMON003 g167961 BLASTN 464 1e−64 81 1827 16 701134363H1SOYMON038 g726031 BLASTN 687 1e−64 84 1828 16 701124677H1 SOYMON037g726031 BLASTN 876 1e−64 85 1829 16 701000359H1 SOYMON018 g1724103BLASTN 877 1e−64 87 1830 16 701139375H1 SOYMON038 g1724103 BLASTN 8831e−64 84 1831 16 700832784H1 SOYMON019 g609224 BLASTN 883 1e−64 89 183216 700980227H1 SOYMON009 g1724103 BLASTN 884 1e−64 84 1833 16700943177H1 SOYMON024 g1655577 BLASTN 885 1e−64 85 1834 16 700844446H1SOYMON021 g2315139 BLASTN 355 1e−63 84 1835 16 700836453H1 SOYMON020g862999 BLASTN 494 1e−63 85 1836 16 700983012H1 SOYMON009 g16508 BLASTN756 1e−63 83 1837 16 700795805H1 SOYMON017 g1655577 BLASTN 833 1e−63 851838 16 701213663H1 SOYMON035 g1724103 BLASTN 862 1e−63 84 1839 16700868366H1 SOYMON016 g167961 BLASTN 864 1e−63 83 1840 16 701134377H1SOYMON038 g2305013 BLASTN 865 1e−63 79 1841 16 LIB3049-007-Q1-E1-C9LIB3049 g16508 BLASTN 867 1e−63 79 1842 16 700944577H1 SOYMON024 g862999BLASTN 869 1e−63 85 1843 16 700992289H1 SOYMON011 g16508 BLASTN 8691e−63 84 1844 16 700891612H1 SOYMON024 g609556 BLASTN 872 1e−63 86 184516 700738277H1 SOYMON012 g609224 BLASTN 463 1e−62 85 1846 16 700895571H1SOYMON027 g609224 BLASTN 562 1e−62 88 1847 16 LIB3049-008-Q1-E1-B3LIB3049 g16508 BLASTN 853 1e−62 77 1848 16 LIB3027-010-Q1-B1-E12 LIB3027g609224 BLASTN 854 1e−62 82 1849 16 701129389H1 SOYMON037 g16508 BLASTN854 1e−62 83 1850 16 701045542H1 SOYMON032 g429105 BLASTN 855 1e−62 861851 16 700969901H1 SOYMON005 g726031 BLASTN 856 1e−62 84 1852 16700984664H1 SOYMON009 g1724103 BLASTN 856 1e−62 81 1853 16 700561352H1SOYMON002 g609224 BLASTN 858 1e−62 85 1854 16 701138828H1 SOYMON038g16508 BLASTN 858 1e−62 84 1855 16 700845962H1 SOYMON021 g1655577 BLASTN861 1e−62 87 1856 16 700896147H1 SOYMON027 g16508 BLASTN 336 1e−61 821857 16 701137929H1 SOYMON038 g1655577 BLASTN 502 1e−61 84 1858 16LIB3040-032-Q1-E1-B8 LIB3040 g16508 BLASTN 521 1e−61 82 1859 16701009537H1 SOYMON019 g726031 BLASTN 545 1e−61 86 1860 16LIB3049-031-Q1-E1-E4 LIB3049 g609224 BLASTN 677 1e−61 82 1861 16LIB3049-031-Q1-E1-C7 LIB3049 g167961 BLASTN 696 1e−61 82 1862 16700891843H1 SOYMON024 g1655577 BLASTN 737 1e−61 84 1863 16 700561231H1SOYMON002 g16508 BLASTN 839 1e−61 82 1864 16 700548238H1 SOYMON002g429105 BLASTN 843 1e−61 85 1865 16 700901018H1 SOYMON027 g2305013BLASTN 845 1e−61 82 1866 16 701134413H1 SOYMON038 g2305013 BLASTN 8481e−61 82 1867 16 700653524H1 SOYMON003 g609224 BLASTN 499 1e−60 85 186816 LIB3049-017-Q1-E1-G8 LIB3049 g16508 BLASTN 514 1e−60 82 1869 16701121077H1 SOYMON037 g16508 BLASTN 671 1e−60 84 1870 16LIB3056-013-Q1-N1-A9 LIB3056 g609224 BLASTN 784 1e−60 86 1871 16700943524H1 SOYMON024 g16508 BLASTN 831 1e−60 85 1872 16 700952889H1SOYMON022 g429103 BLASTN 835 1e−60 84 1873 16 700730632H1 SOYMON009g1655577 BLASTN 431 1e−59 82 1874 16 700649136H1 SOYMON003 g609224BLASTN 486 1e−59 84 1875 16 700895591H1 SOYMON027 g609556 BLASTN 4921e−59 87 1876 16 701009829H1 SOYMON019 g429103 BLASTN 572 1e−59 80 187716 LIB3039-011-Q1-E1-C11 LIB3039 g16508 BLASTN 619 1e−59 83 1878 16LIB3040-055-Q1-E1-C4 LIB3040 g16508 BLASTN 622 1e−59 82 1879 16701123571H1 SOYMON037 g2305013 BLASTN 682 1e−59 81 1880 16LIB3049-004-Q1-E1-E5 LIB3049 g167961 BLASTN 771 1e−59 83 1881 16701002239H1 SOYMON018 g609224 BLASTN 782 1e−59 86 1882 16 700971088H1SOYMON005 g1724103 BLASTN 793 1e−59 81 1883 16 700864624H1 SOYMON016g167961 BLASTN 817 1e−59 83 1884 16 700863534H1 SOYMON027 g16508 BLASTN818 1e−59 86 1885 16 700749489H1 SOYMON013 g16508 BLASTN 823 1e−59 861886 16 700730689H1 SOYMON009 g16508 BLASTN 825 1e−59 83 1887 16LIB3049-050-Q1-E1-A11 LIB3049 g16508 BLASTN 831 1e−59 83 1888 16700850803H1 SOYMON023 g1655577 BLASTN 511 1e−58 83 1889 16 700992317H1SOYMON011 g1655577 BLASTN 520 1e−58 79 1890 16 701119129H1 SOYMON037g16508 BLASTN 553 1e−58 83 1891 16 700657623H1 SOYMON004 g1724103 BLASTN600 1e−58 83 1892 16 LIB3049-015-Q1-E1-F8 LIB3049 g16508 BLASTN 6741e−58 79 1893 16 701007027H1 SOYMON019 g16508 BLASTN 804 1e−58 81 189416 701120820H1 SOYMON037 g609224 BLASTN 807 1e−58 85 1895 16 700654317H1SOYMON004 g1655577 BLASTN 811 1e−58 84 1896 16 700889071H1 SOYMON024g497899 BLASTN 812 1e−58 83 1897 16 701010676H1 SOYMON019 g609224 BLASTN813 1e−58 85 1898 16 701099590H1 SOYMON028 g1655577 BLASTN 486 1e−57 821899 16 700649684H1 SOYMON003 g16508 BLASTN 540 1e−57 82 1900 16700994107H1 SOYMON011 g16508 BLASTN 567 1e−57 80 1901 16 700898629H1SOYMON027 g16508 BLASTN 687 1e−57 82 1902 16 700902333H1 SOYMON027g609224 BLASTN 700 1e−57 82 1903 16 LIB3039-017-Q1-E1-D9 LIB3039 g16508BLASTN 703 1e−57 81 1904 16 LIB3040-026-Q1-E1-A4 LIB3040 g16508 BLASTN709 1e−57 83 1905 16 701130101H1 SOYMON037 g1724103 BLASTN 791 1e−57 881906 16 700902482H1 SOYMON027 g169664 BLASTN 797 1e−57 81 1907 16700730789H1 SOYMON009 g16508 BLASTN 798 1e−57 82 1908 16 700868670H1SOYMON016 g16508 BLASTN 800 1e−57 83 1909 16 LIB3029-011-Q1-B1-D1LIB3029 g609224 BLASTN 800 1e−57 82 1910 16 701119278H1 SOYMON037g609224 BLASTN 801 1e−57 83 1911 16 700747731H1 SOYMON013 g16508 BLASTN801 1e−57 86 1912 16 LIB3049-017-Q1-E1-A11 LIB3049 g16508 BLASTN 8111e−57 80 1913 16 700890428H1 SOYMON024 g609224 BLASTN 482 1e−56 83 191416 700562326H1 SOYMON002 g609224 BLASTN 778 1e−56 82 1915 16 700567740H1SOYMON002 g609224 BLASTN 781 1e−56 85 1916 16 701061514H1 SOYMON033g1724103 BLASTN 784 1e−56 86 1917 16 701135670H1 SOYMON038 g429105BLASTN 785 1e−56 81 1918 16 701139657H1 SOYMON038 g609224 BLASTN 7861e−56 85 1919 16 701123371H1 SOYMON037 g16508 BLASTN 788 1e−56 86 192016 700838744H1 SOYMON020 g16508 BLASTN 789 1e−56 82 1921 16 701070458H1SOYMON034 g1655577 BLASTN 457 1e−55 78 1922 16 701003437H1 SOYMON019g429105 BLASTN 474 1e−55 83 1923 16 700835976H1 SOYMON019 g16508 BLASTN562 1e−55 84 1924 16 700894535H1 SOYMON024 g16508 BLASTN 570 1e−55 861925 16 700752755H1 SOYMON014 g1655577 BLASTN 690 1e−55 83 1926 16LIB3049-032-Q1-E1-C9 LIB3049 g16508 BLASTN 700 1e−55 83 1927 16LIB3040-030-Q1-E1-B5 LIB3040 g16508 BLASTN 709 1e−55 84 1928 16701136935H1 SOYMON038 g609224 BLASTN 766 1e−55 85 1929 16 700682088H1SOYMON008 g169664 BLASTN 767 1e−55 90 1930 16 700682188H1 SOYMON008g169664 BLASTN 767 1e−55 90 1931 16 701068649H1 SOYMON034 g16508 BLASTN770 1e−55 83 1932 16 700726623H1 SOYMON009 g16508 BLASTN 771 1e−55 851933 16 700751129H1 SOYMON014 g16508 BLASTN 772 1e−55 86 1934 16700945234H1 SOYMON024 g16508 BLASTN 773 1e−55 83 1935 16 701133507H2SOYMON038 g609224 BLASTN 776 1e−55 85 1936 16 700902459H1 SOYMON027g726031 BLASTN 777 1e−55 84 1937 16 700986624H1 SOYMON009 g16508 BLASTN612 1e−54 83 1938 16 701097020H1 SOYMON028 g609224 BLASTN 615 1e−54 841939 16 701131409H1 SOYMON038 g16508 BLASTN 755 1e−54 91 1940 16700750123H1 SOYMON013 g726031 BLASTN 756 1e−54 86 1941 16 701040251H1SOYMON029 g16508 BLASTN 756 1e−54 85 1942 16 700732290H1 SOYMON010g169664 BLASTN 758 1e−54 90 1943 16 701117458H1 SOYMON037 g609224 BLASTN759 1e−54 82 1944 16 701118709H1 SOYMON037 g16508 BLASTN 760 1e−54 911945 16 701012834H1 SOYMON019 g16508 BLASTN 760 1e−54 91 1946 16701133316H1 SOYMON038 g16508 BLASTN 760 1e−54 91 1947 16 701129646H1SOYMON037 g16508 BLASTN 760 1e−54 91 1948 16 700732265H1 SOYMON010g169664 BLASTN 760 1e−54 90 1949 16 701040796H1 SOYMON029 g1724103BLASTN 760 1e−54 89 1950 16 700846350H1 SOYMON021 g16508 BLASTN 7611e−54 86 1951 16 701137005H1 SOYMON038 g16508 BLASTN 761 1e−54 86 195216 701046506H1 SOYMON032 g16508 BLASTN 761 1e−54 91 1953 16 700894462H1SOYMON024 g16508 BLASTN 761 1e−54 86 1954 16 700973847H1 SOYMON005g16508 BLASTN 766 1e−54 87 1955 16 701128215H1 SOYMON037 g609556 BLASTN466 1e−53 82 1956 16 700842468H1 SOYMON020 g429105 BLASTN 548 1e−53 861957 16 701051552H1 SOYMON032 g609224 BLASTN 744 1e−53 85 1958 16700944049H1 SOYMON024 g609224 BLASTN 744 1e−53 85 1959 16 701120261H1SOYMON037 g609224 BLASTN 746 1e−53 84 1960 16 700897524H1 SOYMON027g609224 BLASTN 751 1e−53 84 1961 16 LIB3049-042-Q1-E1-F7 LIB3049 g450548BLASTN 761 1e−53 75 1962 16 700896909H1 SOYMON027 g429105 BLASTN 5361e−52 82 1963 16 700974820H1 SOYMON005 g1724103 BLASTN 582 1e−52 86 196416 701143224H1 SOYMON038 g609224 BLASTN 731 1e−52 84 1965 16 701010322H1SOYMON019 g609224 BLASTN 731 1e−52 85 1966 16 701130510H1 SOYMON038g609224 BLASTN 732 1e−52 83 1967 16 700724904H1 SOYMON009 g609224 BLASTN733 1e−52 85 1968 16 701119845H1 SOYMON037 g609224 BLASTN 734 1e−52 851969 16 701107871H1 SOYMON036 g16508 BLASTN 737 1e−52 86 1970 16700983380H1 SOYMON009 g609224 BLASTN 738 1e−52 84 1971 16 700896469H1SOYMON027 g16508 BLASTN 741 1e−52 86 1972 16 700792178H1 SOYMON011g16508 BLASTN 742 1e−52 85 1973 16 701099906H1 SOYMON028 g16508 BLASTN431 1e−51 89 1974 16 701134724H2 SOYMON038 g16508 BLASTN 553 1e−51 851975 16 700749712H1 SOYMON013 g16508 BLASTN 689 1e−51 87 1976 16700982567H1 SOYMON009 g609224 BLASTN 723 1e−51 85 1977 16 701013758H1SOYMON019 g609224 BLASTN 723 1e−51 85 1978 16 700868508H1 SOYMON016g609224 BLASTN 723 1e−51 85 1979 16 700749316H1 SOYMON013 g609224 BLASTN723 1e−51 85 1980 16 701045367H1 SOYMON032 g16508 BLASTN 724 1e−51 911981 16 701205494H1 SOYMON035 g609224 BLASTN 724 1e−51 84 1982 16700556784H1 SOYMON001 g16508 BLASTN 724 1e−51 91 1983 16 701099879H1SOYMON028 g16508 BLASTN 726 1e−51 80 1984 16 701131671H1 SOYMON038g609224 BLASTN 728 1e−51 85 1985 16 701011505H1 SOYMON019 g609224 BLASTN728 1e−51 85 1986 16 701009973H2 SOYMON019 g609224 BLASTN 728 1e−51 851987 16 700793520H1 SOYMON017 g609224 BLASTN 728 1e−51 85 1988 16700753622H1 SOYMON014 g609224 BLASTN 728 1e−51 85 1989 16 700984052H1SOYMON009 g609224 BLASTN 728 1e−51 83 1990 16 701108521H1 SOYMON036g609224 BLASTN 728 1e−51 85 1991 16 700957203H1 SOYMON022 g1724103BLASTN 464 1e−50 86 1992 16 700554120H1 SOYMON001 g609224 BLASTN 5281e−50 84 1993 16 700555954H1 SOYMON001 g167961 BLASTN 528 1e−50 86 199416 701124141H1 SOYMON037 g609224 BLASTN 537 1e−50 85 1995 16 701003232H1SOYMON019 g16508 BLASTN 544 1e−50 88 1996 16 700653112H1 SOYMON003g16508 BLASTN 559 1e−50 91 1997 16 701103353H1 SOYMON028 g1655577 BLASTN581 1e−50 84 1998 16 700562790H1 SOYMON002 g497899 BLASTN 708 1e−50 861999 16 701097303H1 SOYMON028 g609224 BLASTN 711 1e−50 84 2000 16700561195H1 SOYMON002 g609224 BLASTN 712 1e−50 84 2001 16 701121162H1SOYMON037 g16508 BLASTN 712 1e−50 88 2002 16 700981317H1 SOYMON009g16508 BLASTN 714 1e−50 89 2003 16 700981595H1 SOYMON009 g16508 BLASTN716 1e−50 86 2004 16 700994305H1 SOYMON011 g609224 BLASTN 528 1e−49 852005 16 701101565H1 SOYMON028 g429105 BLASTN 552 1e−49 82 2006 16701103456H1 SOYMON028 g429105 BLASTN 617 1e−49 81 2007 16 700993331H1SOYMON011 g16508 BLASTN 622 1e−49 86 2008 16 LIB3052-011-Q1-N1-B12LIB3052 g16508 BLASTN 695 1e−49 85 2009 16 701140613H1 SOYMON038 g609224BLASTN 697 1e−49 85 2010 16 700745426H1 SOYMON013 g16508 BLASTN 6971e−49 84 2011 16 701046530H1 SOYMON032 g609224 BLASTN 697 1e−49 84 201216 701003787H1 SOYMON019 g609224 BLASTN 697 1e−49 85 2013 16 700840830H1SOYMON020 g1724103 BLASTN 699 1e−49 79 2014 16 700901588H1 SOYMON027g609224 BLASTN 702 1e−49 85 2015 16 701037824H1 SOYMON029 g497899 BLASTN702 1e−49 87 2016 16 701131888H1 SOYMON038 g609224 BLASTN 702 1e−49 852017 16 701009947H2 SOYMON019 g609224 BLASTN 702 1e−49 85 2018 16700729216H1 SOYMON009 g497899 BLASTN 702 1e−49 87 2019 16 700831705H1SOYMON019 g609224 BLASTN 702 1e−49 85 2020 16 700900329H1 SOYMON027g429105 BLASTN 705 1e−49 88 2021 16 700563946H1 SOYMON002 g16508 BLASTN705 1e−49 83 2022 16 700808370H1 SOYMON024 g609556 BLASTN 459 1e−48 832023 16 LIB3040-001-Q1-E1-H4 LIB3040 g1724103 BLASTN 475 1e−48 76 202416 700989482H1 SOYMON011 g16508 BLASTN 498 1e−48 86 2025 16 701001311H1SOYMON018 g16508 BLASTN 655 1e−48 85 2026 16 700734733H1 SOYMON010g497899 BLASTN 682 1e−48 87 2027 16 701013070H1 SOYMON019 g497899 BLASTN682 1e−48 87 2028 16 701120989H1 SOYMON037 g16508 BLASTN 683 1e−48 912029 16 LIB3049-025-Q1-E1-B4 LIB3049 g167961 BLASTN 687 1e−48 82 2030 16700808455H1 SOYMON024 g16508 BLASTN 688 1e−48 87 2031 16 701101319H1SOYMON028 g16508 BLASTN 688 1e−48 91 2032 16 701037087H1 SOYMON029g609224 BLASTN 689 1e−48 85 2033 16 701205225H1 SOYMON035 g16508 BLASTN689 1e−48 84 2034 16 701136807H1 SOYMON038 g609224 BLASTN 690 1e−48 852035 16 701118459H1 SOYMON037 g609224 BLASTN 690 1e−48 85 2036 16700942825H1 SOYMON024 g16508 BLASTN 691 1e−48 85 2037 16 701139796H1SOYMON038 g497899 BLASTN 692 1e−48 87 2038 16 700557040H1 SOYMON001g609224 BLASTN 692 1e−48 85 2039 16 701015715H1 SOYMON038 g497899 BLASTN692 1e−48 87 2040 16 700726424H1 SOYMON009 g497899 BLASTN 693 1e−48 872041 16 700790811H1 SOYMON011 g497899 BLASTN 693 1e−48 87 2042 16LIB3040-038-Q1-E1-E5 LIB3040 g16508 BLASTN 709 1e−48 85 2043 16700896626H1 SOYMON027 g429107 BLASTN 430 1e−47 84 2044 16 700562158H1SOYMON002 g609224 BLASTN 518 1e−47 84 2045 16 700747095H1 SOYMON013g16508 BLASTN 538 1e−47 87 2046 16 700726532H1 SOYMON009 g609224 BLASTN622 1e−47 85 2047 16 701137686H1 SOYMON038 g497899 BLASTN 671 1e−47 852048 16 701009148H1 SOYMON019 g16508 BLASTN 673 1e−47 91 2049 16701048279H1 SOYMON032 g16508 BLASTN 674 1e−47 86 2050 16 701120185H1SOYMON037 g16508 BLASTN 674 1e−47 91 2051 16 701040355H1 SOYMON029g497899 BLASTN 675 1e−47 86 2052 16 701130203H1 SOYMON037 g609224 BLASTN678 1e−47 83 2053 16 701015794H1 SOYMON038 g16508 BLASTN 681 1e−47 912054 16 700732181H1 SOYMON010 g16508 BLASTN 681 1e−47 86 2055 16LIB3050-018-Q1-E1-D10 LIB3050 g2305013 BLASTN 696 1e−47 81 2056 16700899157H1 SOYMON027 g609224 BLASTN 486 1e−46 84 2057 16 701119905H1SOYMON037 g497899 BLASTN 507 1e−46 86 2058 16 701062873H1 SOYMON033g497899 BLASTN 509 1e−46 88 2059 16 701210886H1 SOYMON035 g16508 BLASTN540 1e−46 84 2060 16 700893278H1 SOYMON024 g16508 BLASTN 542 1e−46 772061 16 701036970H1 SOYMON029 g16508 BLASTN 579 1e−46 88 2062 16700901932H1 SOYMON027 g609224 BLASTN 640 1e−46 85 2063 16 701044214H1SOYMON032 g429105 BLASTN 658 1e−46 85 2064 16 701056917H1 SOYMON033g16508 BLASTN 661 1e−46 85 2065 16 700981560H1 SOYMON009 g497899 BLASTN661 1e−46 87 2066 16 701213107H1 SOYMON035 g609224 BLASTN 661 1e−46 832067 16 700834235H1 SOYMON019 g497899 BLASTN 661 1e−46 87 2068 16700749413H1 SOYMON013 g16508 BLASTN 662 1e−46 85 2069 16 700889148H1SOYMON024 g16508 BLASTN 662 1e−46 85 2070 16 700745009H1 SOYMON013g609224 BLASTN 662 1e−46 85 2071 16 701206618H1 SOYMON035 g16508 BLASTN663 1e−46 91 2072 16 701131927H1 SOYMON038 g497899 BLASTN 666 1e−46 872073 16 701119069H1 SOYMON037 g497899 BLASTN 666 1e−46 87 2074 16701106908H1 SOYMON036 g497899 BLASTN 666 1e−46 87 2075 16 701103063H1SOYMON028 g497899 BLASTN 666 1e−46 87 2076 16 700686659H1 SOYMON008g166873 BLASTN 667 1e−46 86 2077 16 701040620H1 SOYMON029 g16508 BLASTN669 1e−46 86 2078 16 701012134H1 SOYMON019 g497899 BLASTN 669 1e−46 852079 16 700957508H1 SOYMON022 g1724103 BLASTN 669 1e−46 82 2080 16701008548H1 SOYMON019 g497899 BLASTN 500 1e−45 87 2081 16 700978303H1SOYMON009 g497899 BLASTN 506 1e−45 87 2082 16 701119990H1 SOYMON037g497899 BLASTN 512 1e−45 87 2083 16 700646222H1 SOYMON012 g16508 BLASTN526 1e−45 85 2084 16 701040631H1 SOYMON029 g609556 BLASTN 542 1e−45 872085 16 700894009H1 SOYMON024 g497899 BLASTN 646 1e−45 87 2086 16700908702H1 SOYMON022 g609224 BLASTN 646 1e−45 85 2087 16 700838642H1SOYMON020 g609224 BLASTN 646 1e−45 85 2088 16 700832089H1 SOYMON019g609224 BLASTN 646 1e−45 85 2089 16 700978279H1 SOYMON009 g609224 BLASTN647 1e−45 80 2090 16 700555058H1 SOYMON001 g16508 BLASTN 649 1e−45 842091 16 700906478H1 SOYMON022 g497899 BLASTN 651 1e−45 87 2092 16700897237H1 SOYMON027 g16508 BLASTN 653 1e−45 86 2093 16 701125786H1SOYMON037 g16508 BLASTN 654 1e−45 84 2094 16 700562836H1 SOYMON002g16508 BLASTN 655 1e−45 84 2095 16 700665969H1 SOYMON005 g497899 BLASTN656 1e−45 87 2096 16 701009838H1 SOYMON019 g497899 BLASTN 656 1e−45 872097 16 701211770H1 SOYMON035 g16508 BLASTN 657 1e−45 84 2098 16700654550H1 SOYMON004 g16508 BLASTN 657 1e−45 83 2099 16 700567949H1SOYMON002 g16508 BLASTN 657 1e−45 84 2100 16 701048175H1 SOYMON032g16508 BLASTN 658 1e−45 91 2101 16 700901037H1 SOYMON027 g16508 BLASTN658 1e−45 86 2102 16 700656917H1 SOYMON004 g1724103 BLASTN 407 1e−44 812103 16 700565684H1 SOYMON002 g16508 BLASTN 463 1e−44 86 2104 16701002808H1 SOYMON019 g497899 BLASTN 503 1e−44 87 2105 16 701107115H1SOYMON036 g497899 BLASTN 507 1e−44 87 2106 16 701122669H1 SOYMON037g16508 BLASTN 636 1e−44 82 2107 16 700894409H1 SOYMON024 g497899 BLASTN636 1e−44 87 2108 16 700848374H1 SOYMON021 g497899 BLASTN 636 1e−44 872109 16 700902420H1 SOYMON027 g497899 BLASTN 636 1e−44 87 2110 16700747450H1 SOYMON013 g16508 BLASTN 638 1e−44 91 2111 16 701125869H1SOYMON037 g16508 BLASTN 638 1e−44 91 2112 16 700731302H1 SOYMON010g16508 BLASTN 639 1e−44 85 2113 16 701099068H1 SOYMON028 g2305013 BLASTN640 1e−44 81 2114 16 700975556H1 SOYMON009 g450548 BLASTN 641 1e−44 842115 16 700889660H1 SOYMON024 g16508 BLASTN 642 1e−44 86 2116 16700951744H1 SOYMON022 g16508 BLASTN 642 1e−44 86 2117 16 700750251H1SOYMON013 g609224 BLASTN 643 1e−44 86 2118 16 700834611H1 SOYMON019g16508 BLASTN 643 1e−44 91 2119 16 700565356H1 SOYMON002 g16508 BLASTN643 1e−44 80 2120 16 700673803H1 SOYMON007 g16508 BLASTN 644 1e−44 842121 16 700958721H1 SOYMON022 g16508 BLASTN 645 1e−44 86 2122 16700985717H1 SOYMON009 g16508 BLASTN 646 1e−44 82 2123 16LIB3049-032-Q1-E1-A5 LIB3049 g16508 BLASTN 662 1e−44 76 2124 16700733454H1 SOYMON010 g609224 BLASTN 432 1e−43 85 2125 16 700987656H1SOYMON009 g497899 BLASTN 476 1e−43 87 2126 16 700755957H1 SOYMON014g497899 BLASTN 483 1e−43 86 2127 16 700749331H1 SOYMON013 g2305013BLASTN 521 1e−43 84 2128 16 700830984H1 SOYMON019 g497899 BLASTN 6221e−43 86 2129 16 700906176H1 SOYMON022 g16508 BLASTN 623 1e−43 86 213016 701100070H2 SOYMON028 g16508 BLASTN 623 1e−43 91 2131 16 700954053H1SOYMON022 g497899 BLASTN 624 1e−43 88 2132 16 700833949H1 SOYMON019g497899 BLASTN 624 1e−43 88 2133 16 700976710H1 SOYMON009 g2305013BLASTN 625 1e−43 79 2134 16 701117925H2 SOYMON037 g16508 BLASTN 6251e−43 84 2135 16 700748825H1 SOYMON013 g16508 BLASTN 626 1e−43 84 213616 700899651H1 SOYMON027 g16508 BLASTN 626 1e−43 84 2137 16 700746739H1SOYMON013 g497899 BLASTN 626 1e−43 88 2138 16 701009051H1 SOYMON019g16508 BLASTN 628 1e−43 91 2139 16 700754423H1 SOYMON014 g16508 BLASTN628 1e−43 91 2140 16 700760701H1 SOYMON015 g16508 BLASTN 628 1e−43 832141 16 701098124H1 SOYMON028 g2305013 BLASTN 628 1e−43 78 2142 16700984979H1 SOYMON009 g16508 BLASTN 628 1e−43 91 2143 16 700565262H1SOYMON002 g497899 BLASTN 629 1e−43 86 2144 16 700954979H1 SOYMON022g497899 BLASTN 630 1e−43 87 2145 16 700834601H1 SOYMON019 g497899 BLASTN630 1e−43 87 2146 16 700953156H1 SOYMON022 g609224 BLASTN 630 1e−43 852147 16 700865217H1 SOYMON016 g16508 BLASTN 631 1e−43 77 2148 16700852947H1 SOYMON023 g16508 BLASTN 631 1e−43 84 2149 16 700943547H1SOYMON024 g497899 BLASTN 631 1e−43 86 2150 16 701139277H1 SOYMON038g609224 BLASTN 632 1e−43 82 2151 16 700900155H1 SOYMON027 g429105 BLASTN264 1e−42 81 2152 16 700970581H1 SOYMON005 g497899 BLASTN 296 1e−42 862153 16 700654232H1 SOYMON003 g497899 BLASTN 348 1e−42 88 2154 16700743608H1 SOYMON012 g497899 BLASTN 354 1e−42 86 2155 16 700986620H1SOYMON009 g609224 BLASTN 360 1e−42 84 2156 16 700562590H1 SOYMON002g16508 BLASTN 476 1e−42 83 2157 16 701119690H1 SOYMON037 g167961 BLASTN506 1e−42 84 2158 16 700951730H1 SOYMON022 g497899 BLASTN 507 1e−42 872159 16 701004525H1 SOYMON019 g16508 BLASTN 516 1e−42 91 2160 16700892637H1 SOYMON024 g16508 BLASTN 523 1e−42 81 2161 16 700560679H1SOYMON001 g16508 BLASTN 526 1e−42 83 2162 16 700897888H1 SOYMON027g16508 BLASTN 611 1e−42 86 2163 16 700547973H1 SOYMON001 g497899 BLASTN611 1e−42 84 2164 16 701012166H1 SOYMON019 g497899 BLASTN 612 1e−42 872165 16 700747550H1 SOYMON013 g16508 BLASTN 613 1e−42 84 2166 16700946136H1 SOYMON024 g16508 BLASTN 615 1e−42 85 2167 16 700665932H1SOYMON005 g16508 BLASTN 616 1e−42 84 2168 16 701010969H1 SOYMON019g16508 BLASTN 618 1e−42 84 2169 16 700962515H1 SOYMON022 g16508 BLASTN618 1e−42 91 2170 16 700895685H1 SOYMON027 g429107 BLASTN 618 1e−42 772171 16 700752015H1 SOYMON014 g16508 BLASTN 620 1e−42 84 2172 16701015704H1 SOYMON038 g16508 BLASTN 620 1e−42 85 2173 16 700966710H1SOYMON028 g16508 BLASTN 620 1e−42 83 2174 16 701004268H1 SOYMON019g16508 BLASTN 620 1e−42 86 2175 16 701138901H1 SOYMON038 g497899 BLASTN285 1e−41 85 2176 16 700648891H1 SOYMON003 g16508 BLASTN 481 1e−41 842177 16 700741421H1 SOYMON012 g609224 BLASTN 511 1e−41 82 2178 16700555653H1 SOYMON001 g166873 BLASTN 512 1e−41 85 2179 16 700958314H1SOYMON022 g497899 BLASTN 599 1e−41 87 2180 16 700960055H1 SOYMON022g497899 BLASTN 599 1e−41 87 2181 16 700901925H1 SOYMON027 g16508 BLASTN599 1e−41 84 2182 16 700741987H1 SOYMON012 g497899 BLASTN 599 1e−41 872183 16 700662837H1 SOYMON005 g497899 BLASTN 599 1e−41 87 2184 16701060927H1 SOYMON033 g497899 BLASTN 599 1e−41 87 2185 16 700562973H1SOYMON002 g16508 BLASTN 601 1e−41 87 2186 16 700754586H1 SOYMON014g16508 BLASTN 603 1e−41 84 2187 16 701132674H1 SOYMON038 g16508 BLASTN604 1e−41 85 2188 16 701010012H2 SOYMON019 g609224 BLASTN 604 1e−41 842189 16 701123643H1 SOYMON037 g16508 BLASTN 604 1e−41 85 2190 16700894196H1 SOYMON024 g16508 BLASTN 604 1e−41 85 2191 16 700899248H1SOYMON027 g16508 BLASTN 604 1e−41 85 2192 16 701207680H1 SOYMON035g16508 BLASTN 605 1e−41 84 2193 16 700899210H1 SOYMON027 g16508 BLASTN605 1e−41 84 2194 16 700898762H1 SOYMON027 g16508 BLASTN 605 1e−41 872195 16 700983636H1 SOYMON009 g497899 BLASTN 606 1e−41 87 2196 16700845511H1 SOYMON021 g16508 BLASTN 606 1e−41 89 2197 16 700563416H1SOYMON002 g16508 BLASTN 606 1e−41 83 2198 16 701004041H1 SOYMON019g16508 BLASTN 607 1e−41 84 2199 16 701097184H1 SOYMON028 g16508 BLASTN607 1e−41 87 2200 16 700750691H1 SOYMON014 g609224 BLASTN 608 1e−41 842201 16 701208091H1 SOYMON035 g16508 BLASTN 608 1e−41 84 2202 16700973888H1 SOYMON005 g609224 BLASTN 609 1e−41 85 2203 16 700831972H1SOYMON019 g16508 BLASTN 609 1e−41 85 2204 16 701010090H2 SOYMON019g16508 BLASTN 610 1e−41 87 2205 16 700890067H1 SOYMON024 g16508 BLASTN610 1e−41 87 2206 16 700833227H1 SOYMON019 g16508 BLASTN 610 1e−41 852207 16 700563171H1 SOYMON002 g16508 BLASTN 370 1e−40 90 2208 16701121457H1 SOYMON037 g609224 BLASTN 372 1e−40 85 2209 16 700566851H1SOYMON002 g16508 BLASTN 402 1e−40 82 2210 16 700753221H1 SOYMON014g609224 BLASTN 422 1e−40 84 2211 16 700749039H1 SOYMON013 g16508 BLASTN475 1e−40 84 2212 16 700558940H1 SOYMON001 g16508 BLASTN 493 1e−40 802213 16 700757695H1 SOYMON015 g16508 BLASTN 526 1e−40 83 2214 16701102262H1 SOYMON028 g2305013 BLASTN 587 1e−40 81 2215 16 700747484H1SOYMON013 g16508 BLASTN 587 1e−40 85 2216 16 700891875H1 SOYMON024g16508 BLASTN 587 1e−40 85 2217 16 700894439H1 SOYMON024 g497899 BLASTN588 1e−40 88 2218 16 701062577H1 SOYMON033 g16508 BLASTN 588 1e−40 872219 16 700964366H1 SOYMON022 g16508 BLASTN 589 1e−40 85 2220 16701210016H1 SOYMON035 g16508 BLASTN 589 1e−40 83 2221 16 700901573H1SOYMON027 g16508 BLASTN 589 1e−40 85 2222 16 700750533H1 SOYMON014g16508 BLASTN 589 1e−40 85 2223 16 700888820H1 SOYMON024 g497899 BLASTN589 1e−40 87 2224 16 700891965H1 SOYMON024 g497899 BLASTN 589 1e−40 872225 16 700865001H1 SOYMON016 g16508 BLASTN 589 1e−40 85 2226 16700654923H1 SOYMON004 g16508 BLASTN 590 1e−40 84 2227 16 700746169H1SOYMON013 g16508 BLASTN 590 1e−40 84 2228 16 700745792H1 SOYMON013g16508 BLASTN 591 1e−40 85 2229 16 700966953H1 SOYMON029 g16508 BLASTN591 1e−40 85 2230 16 701141657H1 SOYMON038 g16508 BLASTN 592 1e−40 862231 16 700943078H1 SOYMON024 g16508 BLASTN 598 1e−40 85 2232 16701137073H1 SOYMON038 g16508 BLASTN 598 1e−40 90 2233 16 700966730H1SOYMON028 g16508 BLASTN 598 1e−40 84 2234 16 701137731H1 SOYMON038g16508 BLASTN 371 1e−39 84 2235 16 701044050H1 SOYMON032 g497899 BLASTN428 1e−39 89 2236 16 700993309H1 SOYMON011 g497899 BLASTN 434 1e−39 882237 16 700874483H1 SOYMON018 g497899 BLASTN 434 1e−39 87 2238 16701014866H1 SOYMON019 g16508 BLASTN 454 1e−39 89 2239 16 700894603H1SOYMON024 g16508 BLASTN 454 1e−39 85 2240 16 700958677H1 SOYMON022g497899 BLASTN 455 1e−39 87 2241 16 700754001H1 SOYMON014 g609224 BLASTN518 1e−39 85 2242 16 700946430H1 SOYMON024 g497899 BLASTN 578 1e−39 872243 16 700729892H1 SOYMON009 g497899 BLASTN 578 1e−39 87 2244 16700753230H1 SOYMON014 g497899 BLASTN 578 1e−39 87 2245 16 700562584H1SOYMON002 g16508 BLASTN 578 1e−39 82 2246 16 700842880H1 SOYMON020g16508 BLASTN 580 1e−39 82 2247 16 700852561H1 SOYMON023 g16508 BLASTN582 1e−39 85 2248 16 700941290H1 SOYMON024 g497899 BLASTN 583 1e−39 872249 16 700750361H1 SOYMON013 g497899 BLASTN 584 1e−39 80 2250 16700841915H1 SOYMON020 g16508 BLASTN 585 1e−39 85 2251 16 700944462H1SOYMON024 g16508 BLASTN 585 1e−39 85 2252 16 700548174H1 SOYMON002g16508 BLASTN 342 1e−38 82 2253 16 700942324H1 SOYMON024 g497899 BLASTN403 1e−38 87 2254 16 700788450H1 SOYMON011 g497899 BLASTN 416 1e−38 862255 16 701127954H1 SOYMON037 g16508 BLASTN 449 1e−38 76 2256 16700742223H1 SOYMON012 g1724103 BLASTN 451 1e−38 77 2257 16 701102722H1SOYMON028 g16508 BLASTN 466 1e−38 92 2258 16 701046957H1 SOYMON032g16508 BLASTN 517 1e−38 85 2259 16 700836169H1 SOYMON019 g497899 BLASTN517 1e−38 87 2260 16 700697967H1 SOYMON015 g16508 BLASTN 521 1e−38 842261 16 700678867H1 SOYMON007 g609224 BLASTN 566 1e−38 82 2262 16700838753H1 SOYMON020 g16508 BLASTN 567 1e−38 85 2263 16 700726468H1SOYMON009 g16508 BLASTN 570 1e−38 85 2264 16 701014631H1 SOYMON019g497899 BLASTN 571 1e−38 87 2265 16 700728719H1 SOYMON009 g497899 BLASTN573 1e−38 87 2266 16 700962582H1 SOYMON022 g497899 BLASTN 573 1e−38 872267 16 701123183H1 SOYMON037 g16508 BLASTN 573 1e−38 84 2268 16700752347H1 SOYMON014 g16508 BLASTN 574 1e−38 87 2269 16 701103315H1SOYMON028 g16508 BLASTN 394 1e−37 86 2270 16 700750955H1 SOYMON014g497899 BLASTN 414 1e−37 88 2271 16 700867493H1 SOYMON016 g609224 BLASTN433 1e−37 85 2272 16 701054760H1 SOYMON032 g16508 BLASTN 475 1e−37 852273 16 700900903H1 SOYMON027 g16508 BLASTN 501 1e−37 84 2274 16701102294H1 SOYMON028 g16508 BLASTN 502 1e−37 87 2275 16 701124034H1SOYMON037 g16508 BLASTN 512 1e−37 82 2276 16 700981476H1 SOYMON009g16508 BLASTN 526 1e−37 83 2277 16 700903712H1 SOYMON022 g609224 BLASTN552 1e−37 85 2278 16 700648751H1 SOYMON003 g1655577 BLASTN 554 1e−37 862279 16 700829930H1 SOYMON019 g609224 BLASTN 556 1e−37 84 2280 16700758078H1 SOYMON015 g16508 BLASTN 556 1e−37 84 2281 16 700754248H1SOYMON014 g16508 BLASTN 557 1e−37 81 2282 16 700834650H1 SOYMON019g609224 BLASTN 557 1e−37 85 2283 16 700746279H1 SOYMON013 g16508 BLASTN558 1e−37 82 2284 16 700869124H1 SOYMON016 g16508 BLASTN 559 1e−37 832285 16 701100219H1 SOYMON028 g2305013 BLASTN 560 1e−37 80 2286 16700992589H1 SOYMON011 g497899 BLASTN 560 1e−37 80 2287 16 700865642H1SOYMON016 g16508 BLASTN 562 1e−37 84 2288 16 701068835H1 SOYMON034g429105 BLASTN 321 1e−36 86 2289 16 700746908H1 SOYMON013 g16508 BLASTN373 1e−36 81 2290 16 700954719H1 SOYMON022 g16508 BLASTN 475 1e−36 862291 16 701042790H1 SOYMON029 g497899 BLASTN 489 1e−36 88 2292 16700835259H1 SOYMON019 g497899 BLASTN 507 1e−36 88 2293 16 700565816H1SOYMON002 g16508 BLASTN 519 1e−36 82 2294 16 700895811H1 SOYMON027g609224 BLASTN 540 1e−36 83 2295 16 700975709H1 SOYMON009 g609224 BLASTN541 1e−36 84 2296 16 700751645H1 SOYMON014 g16508 BLASTN 542 1e−36 852297 16 700567087H1 SOYMON002 g16508 BLASTN 542 1e−36 85 2298 16700893027H1 SOYMON024 g609224 BLASTN 542 1e−36 85 2299 16 700891185H1SOYMON024 g609224 BLASTN 543 1e−36 84 2300 16 700851376H1 SOYMON023g16508 BLASTN 543 1e−36 83 2301 16 700898322H1 SOYMON027 g609224 BLASTN547 1e−36 85 2302 16 700762881H1 SOYMON015 g16508 BLASTN 323 1e−35 852303 16 700960023H1 SOYMON022 g16508 BLASTN 370 1e−35 90 2304 16701136486H1 SOYMON038 g497899 BLASTN 378 1e−35 85 2305 16 700987688H1SOYMON009 g497899 BLASTN 387 1e−35 83 2306 16 700836508H1 SOYMON020g16508 BLASTN 431 1e−35 85 2307 16 700990779H1 SOYMON011 g167961 BLASTN471 1e−35 86 2308 16 700753019H1 SOYMON014 g497899 BLASTN 476 1e−35 832309 16 701138490H1 SOYMON038 g16508 BLASTN 481 1e−35 83 2310 16700751771H1 SOYMON014 g609224 BLASTN 528 1e−35 84 2311 16 701048324H1SOYMON032 g609224 BLASTN 529 1e−35 84 2312 16 700835739H1 SOYMON019g16508 BLASTN 532 1e−35 85 2313 16 700986295H1 SOYMON009 g497899 BLASTN533 1e−35 86 2314 16 701205693H1 SOYMON035 g16508 BLASTN 535 1e−35 842315 16 700865674H1 SOYMON016 g609224 BLASTN 536 1e−35 85 2316 16700943777H1 SOYMON024 g16508 BLASTN 536 1e−35 84 2317 16 700968207H1SOYMON035 g609224 BLASTN 536 1e−35 85 2318 16 700898370H1 SOYMON027g609224 BLASTN 536 1e−35 85 2319 16 700987890H1 SOYMON009 g609224 BLASTN536 1e−35 85 2320 16 700896016H1 SOYMON027 g609224 BLASTN 536 1e−35 852321 16 700554440H1 SOYMON001 g16508 BLASTN 283 1e−34 82 2322 16700789855H2 SOYMON011 g16508 BLASTN 293 1e−34 91 2323 16 701101393H1SOYMON028 g16508 BLASTN 311 1e−34 84 2324 16 700946335H1 SOYMON024g16508 BLASTN 323 1e−34 86 2325 16 701207420H1 SOYMON035 g169664 BLASTN350 1e−34 88 2326 16 700733034H1 SOYMON010 g16508 BLASTN 384 1e−34 812327 16 700790653H2 SOYMON011 g497899 BLASTN 399 1e−34 81 2328 16700901808H1 SOYMON027 g497899 BLASTN 430 1e−34 81 2329 16 700760954H1SOYMON015 g609224 BLASTN 514 1e−34 84 2330 16 700896206H1 SOYMON027g16508 BLASTN 516 1e−34 85 2331 16 701039157H1 SOYMON029 g609224 BLASTN520 1e−34 76 2332 16 700991056H1 SOYMON011 g166873 BLASTN 520 1e−34 802333 16 701097173H1 SOYMON028 g609224 BLASTN 521 1e−34 85 2334 16700900989H1 SOYMON027 g497899 BLASTN 521 1e−34 86 2335 16 700895584H1SOYMON027 g609224 BLASTN 521 1e−34 85 2336 16 700726668H1 SOYMON009g16508 BLASTN 521 1e−34 85 2337 16 701011191H1 SOYMON019 g16508 BLASTN521 1e−34 85 2338 16 701100827H1 SOYMON028 g16508 BLASTN 522 1e−34 842339 16 701156732H1 SOYMON031 g16508 BLASTN 522 1e−34 84 2340 16700889752H1 SOYMON024 g609224 BLASTN 523 1e−34 84 2341 16 701105873H1SOYMON036 g16508 BLASTN 524 1e−34 87 2342 16 700958346H1 SOYMON022g497899 BLASTN 525 1e−34 84 2343 16 700962972H1 SOYMON022 g609224 BLASTN525 1e−34 84 2344 16 701055918H1 SOYMON032 g16508 BLASTN 526 1e−34 852345 16 701123061H1 SOYMON037 g16508 BLASTN 526 1e−34 85 2346 16701109796H1 SOYMON036 g166873 BLASTN 335 1e−33 86 2347 16 701207350H1SOYMON035 g497899 BLASTN 349 1e−33 89 2348 16 700835692H1 SOYMON019g16508 BLASTN 377 1e−33 81 2349 16 700729083H1 SOYMON009 g16508 BLASTN448 1e−33 85 2350 16 LIB3040-049-Q1-E1-E8 LIB3040 g16508 BLASTN 4641e−33 80 2351 16 700896567H1 SOYMON027 g497899 BLASTN 502 1e−33 81 235216 700668032H1 SOYMON006 g609224 BLASTN 504 1e−33 85 2353 16 700895062H1SOYMON024 g609224 BLASTN 505 1e−33 83 2354 16 700961178H1 SOYMON022g16508 BLASTN 506 1e−33 84 2355 16 701202590H1 SOYMON035 g497899 BLASTN508 1e−33 86 2356 16 700754960H1 SOYMON014 g16508 BLASTN 512 1e−33 832357 16 700748584H1 SOYMON013 g16508 BLASTN 512 1e−33 84 2358 16700791669H1 SOYMON011 g609224 BLASTN 512 1e−33 84 2359 16 701210405H1SOYMON035 g16508 BLASTN 526 1e−33 83 2360 16 LIB3040-054-Q1-E1-D8LIB3040 g16508 BLASTN 531 1e−33 80 2361 16 700791176H1 SOYMON011 g497899BLASTN 311 1e−32 87 2362 16 LIB3049-029-Q1-E1-C5 LIB3049 g167961 BLASTN368 1e−32 73 2363 16 700893458H1 SOYMON024 g497899 BLASTN 380 1e−32 872364 16 700829728H1 SOYMON019 g497899 BLASTN 490 1e−32 88 2365 16700833137H1 SOYMON019 g609224 BLASTN 490 1e−32 85 2366 16 701204592H2SOYMON035 g609224 BLASTN 493 1e−32 86 2367 16 700834821H1 SOYMON019g609224 BLASTN 494 1e−32 78 2368 16 700757167H1 SOYMON015 g16508 BLASTN494 1e−32 85 2369 16 701203730H2 SOYMON035 g16508 BLASTN 496 1e−32 822370 16 701044359H1 SOYMON032 g609224 BLASTN 497 1e−32 84 2371 16701010095H2 SOYMON019 g16508 BLASTN 498 1e−32 86 2372 16 701100059H2SOYMON028 g16508 BLASTN 499 1e−32 80 2373 16 700844256H1 SOYMON021g16508 BLASTN 500 1e−32 80 2374 16 700968296H1 SOYMON035 g497899 BLASTN501 1e−32 85 2375 16 701110404H1 SOYMON036 g16508 BLASTN 333 1e−31 842376 16 700869025H1 SOYMON016 g497899 BLASTN 340 1e−31 86 2377 16700556245H1 SOYMON001 g609224 BLASTN 355 1e−31 84 2378 16 700566166H1SOYMON002 g726031 BLASTN 402 1e−31 77 2379 16 700941481H1 SOYMON024g497899 BLASTN 423 1e−31 79 2380 16 700896940H1 SOYMON027 g169664 BLASTN477 1e−31 85 2381 16 701145429H1 SOYMON031 g609224 BLASTN 478 1e−31 852382 16 700946496H1 SOYMON024 g609224 BLASTN 488 1e−31 86 2383 16700902149H1 SOYMON027 g1655577 BLASTN 488 1e−31 87 2384 16 700653072H1SOYMON003 g497899 BLASTN 489 1e−31 88 2385 16 701000261H1 SOYMON018g16508 BLASTN 495 1e−31 78 2386 16 701039358H1 SOYMON029 g609224 BLASTN320 1e−30 85 2387 16 701156951H1 SOYMON031 g609224 BLASTN 467 1e−30 862388 16 701040880H1 SOYMON029 g497899 BLASTN 477 1e−30 89 2389 16700979632H2 SQYMON009 g609224 BLASTN 490 1e−30 83 2390 16 700983678H1SOYMON009 g16508 BLASTN 192 1e−29 86 2391 16 700893644H1 SOYMON024g609224 BLASTN 320 1e−29 85 2392 16 700962437H1 SOYMON022 g609224 BLASTN364 1e−29 83 2393 16 701150545H1 SOYMON031 g16508 BLASTN 457 1e−29 862394 16 700555403H1 SOYMON001 g16508 BLASTN 458 1e−29 90 2395 16700794307H1 SOYMON017 g16508 BLASTN 460 1e−29 87 2396 16 701137971H1SOYMON038 g16508 BLASTN 460 1e−29 84 2397 16 700893774H1 SOYMON024g609224 BLASTN 461 1e−29 86 2398 16 700565272H1 SOYMON002 g609224 BLASTN464 1e−29 74 2399 16 700667161H1 SOYMON006 g16508 BLASTN 468 1e−29 752400 16 700752952H1 SOYMON014 g609224 BLASTN 279 1e−28 78 2401 16700901534H1 SOYMON027 g2305013 BLASTN 288 1e−28 85 2402 16 701006617H1SOYMON019 g497899 BLASTN 358 1e−28 79 2403 16 700753459H1 SOYMON014g609224 BLASTN 446 1e−28 81 2404 16 701157089H1 SOYMON031 g609224 BLASTN448 1e−28 84 2405 16 700831271H1 SOYMON019 g16508 BLASTN 453 1e−28 872406 16 701207325H1 SOYMON035 g167961 BLASTN 251 1e−27 81 2407 16700566121H1 SOYMON002 g16508 BLASTN 259 1e−27 75 2408 16 701137878H1SOYMON038 g609224 BLASTN 280 1e−27 85 2409 16 700763957H1 SOYMON019g497899 BLASTN 430 1e−27 88 2410 16 701015858H1 SOYMON038 g609224 BLASTN431 1e−27 86 2411 16 700742849H1 SOYMON012 g497899 BLASTN 434 1e−27 882412 16 701102648H1 SOYMON028 g16508 BLASTN 436 1e−27 89 2413 16700940955H1 SOYMON024 g609556 BLASTN 438 1e−27 85 2414 16 700889805H1SOYMON024 g16508 BLASTN 441 1e−27 89 2415 16 701061930H1 SOYMON033g16508 BLASTN 315 1e−26 77 2416 16 700901375H1 SOYMON027 g609224 BLASTN323 1e−26 84 2417 16 700989056H1 SOYMON011 g609224 BLASTN 345 1e−26 822418 16 700665988H1 SOYMON005 g960356 BLASTN 420 1e−26 79 2419 16700893352H1 SOYMON024 g497899 BLASTN 428 1e−26 87 2420 16 701122945H1SOYMON037 g16508 BLASTN 269 1e−25 76 2421 16 700992929H1 SOYMON011g16508 BLASTN 328 1e−25 78 2422 16 700557564H1 SOYMON001 g609224 BLASTN413 1e−25 85 2423 16 701102620H1 SOYMON028 g609224 BLASTN 414 1e−25 822424 16 701062321H1 SOYMON033 g497899 BLASTN 267 1e−24 87 2425 16701062258H1 SOYMON033 g450548 BLASTN 391 1e−24 83 2426 16 701145703H1SOYMON031 g609224 BLASTN 411 1e−24 85 2427 16 701144943H1 SOYMON031g450548 BLASTN 421 1e−24 85 2428 16 701134190H1 SOYMON038 g2305013BLASTN 268 1e−23 87 2429 16 701132116H1 SOYMON038 g450548 BLASTN 3851e−23 85 2430 16 701144522H1 SOYMON031 g450548 BLASTN 406 1e−23 85 243116 700738002H1 SOYMON012 g1655578 BLASTX 205 1e−21 84 2432 16700756686H1 SOYMON014 g16508 BLASTN 235 1e−21 76 2433 16 700649058H1SOYMON003 g16508 BLASTN 360 1e−21 85 2434 16 701152050H1 SOYMON031g497899 BLASTN 233 1e−20 87 2435 16 701156991H1 SOYMON031 g497900 BLASTX146 1e−19 98 2436 16 700761949H1 SOYMON015 g497900 BLASTX 173 1e−19 982437 16 700988163H1 SOYMON009 g609224 BLASTN 243 1e−19 81 2438 16700666050H1 SOYMON005 g960357 BLASTX 183 1e−18 89 2439 16 700896896H1SOYMON027 g497900 BLASTX 95 1e−17 82 2440 16 701100249H1 SOYMON028g497900 BLASTX 104 1e−17 79 2441 16 701061719H1 SOYMON033 g609224 BLASTN310 1e−17 67 2442 16 700674836H1 SOYMON007 g16508 BLASTN 331 1e−17 842443 16 700908822H1 SOYMON022 g169665 BLASTX 171 1e−16 100 2444 16700898670H1 SOYMON027 g166872 BLASTX 172 1e−16 93 2445 16 700650967H1SOYMON003 g16845 BLASTX 147 1e−15 100 2446 16 701132185H1 SOYMON038g16845 BLASTX 151 1e−15 100 2447 16 700648929H1 SOYMON003 g1033190BLASTX 156 1e−14 96 2448 16 700960809H1 SOYMON022 g16508 BLASTN 1681e−14 88 2449 16 701101753H1 SOYMON028 g16845 BLASTX 141 1e−13 88 245016 700742010H1 SOYMON012 g609224 BLASTN 273 1e−13 88 2451 16 700735080H1SOYMON010 g166874 BLASTX 121 1e−12 86 2452 16 700979242H1 SOYMON009g609224 BLASTN 165 1e−12 81 2453 16 700740838H1 SOYMON012 g609224 BLASTN253 1e−12 88 2454 16 700832383H1 SOYMON019 g16845 BLASTX 82 1e−11 712455 16 700830938H1 SOYMON019 g16845 BLASTX 92 1e−11 90 2456 16700564686H1 SOYMON002 g1655578 BLASTX 95 1e−11 97 2457 16 700753178H1SOYMON014 g16845 BLASTX 107 1e−10 78 2458 16 700563834H1 SOYMON002g497900 BLASTX 124 1e−10 100 2459 16 700897739H1 SOYMON027 g16845 BLASTX125 1e−10 86 2460 16 700725722H1 SOYMON009 g609225 BLASTX 125 1e−10 922461 16 701210357H1 SOYMON035 g497900 BLASTX 128 1e−10 100 2462 16700833654H1 SOYMON019 g609224 BLASTN 243 1e−10 87 2463 16 701010334H1SOYMON019 g609224 BLASTN 258 1e−10 86 2464 16 700567622H1 SOYMON002g166874 BLASTX 93 1e−9 100 2465 16 700647933H1 SOYMON003 g609225 BLASTX120 1e−9 81 2466 16 700742255H1 SOYMON012 g429107 BLASTN 160 1e−9 862467 16 700981506H1 SOYMON009 g609224 BLASTN 248 1e−9 88 2468 16700962044H1 SOYMON022 g497900 BLASTX 113 1e−8 100 2469 16 701039604H1SOYMON029 g497900 BLASTX 113 1e−8 100 2470 16 701009941H2 SOYMON019g609225 BLASTX 114 1e−8 70 2471 16 700976956H1 SOYMON009 g609225 BLASTX118 1e−8 96 2472 18138 701120722H1 SOYMON037 g169664 BLASTN 514 1e−33 922473 18138 700946044H1 SOYMON024 g169664 BLASTN 437 1e−26 90 2474 18138700664443H1 SOYMON005 g17262 BLASTX 162 1e−16 88 2475 18138 700665162H1SOYMON005 g17262 BLASTX 162 1e−16 88 2476 18138 701143667H1 SOYMON038g16961 BLASTX 123 1e−11 95 2477 18138 701099150H1 SOYMON028 g16961BLASTX 112 1e−9 90 2478 27686 700909141H1 SOYMON022 g726027 BLASTN 7231e−51 84 2479 27686 701145458H1 SOYMON031 g726027 BLASTN 694 1e−49 86

ADENOSYLMETHIONINE DECARBOXYLASE (EC 4.1.1.50) Seq No. Cluster IDCloneID Library NCBI gi Method Score P-value % Ident 430 -700151703700151703H1 SATMON007 g1532072 BLASTN 717 1e−50 91 431 -700165608700165608H1 SATMON013 g1532072 BLASTN 605 1e−41 75 432 -700166868700166868H1 SATMON013 g1532072 BLASTN 593 1e−40 90 433 -700242148700242148H1 SATMON010 g1532048 BLASTX 142 1e−12 88 434 -700354923700354923H1 SATMON024 g1532072 BLASTN 887 1e−74 89 435 -700422279700422279H1 SATMONN01 g1532072 BLASTN 623 1e−61 88 436 -700455682700455682H1 SATMON029 g1532047 BLASTN 681 1e−47 77 437 -700477645700477645H1 SATMON025 g1532048 BLASTX 90 1e−12 71 438 -700550509700550509H1 SATMON022 g1532047 BLASTN 399 1e−41 80 439 -700572502700572502H1 SATMON030 g1532048 BLASTX 93 1e−20 70 440 -700618258700618258H1 SATMON033 g1532072 BLASTN 368 1e−67 92 441 -701165456701165456H1 SATMONN04 g1532072 BLASTN 436 1e−26 71 442 -L30622289LIB3062-004-Q1-K1-E10 LIB3062 g1532047 BLASTN 406 1e−26 82 443-L30623185 LIB3062-026-Q1-K1-E2 LIB3062 g1532072 BLASTN 338 1e−19 62 444-L30625601 LIB3062-033-Q1-K1-A9 LIB3062 g1403043 BLASTN 758 1e−54 68 445-L30661842 LIB3066-009-Q1-K1-E4 LIB3066 g1532072 BLASTN 398 1e−22 86 446-L30681932 LIB3068-020-Q1-K1-C8 LIB3068 g1532072 BLASTN 205 1e−28 89 447-L30687176 LIB3068-059-Q1-K1-C1 LIB3068 g1532072 BLASTN 216 1e−20 87 448-L30692075 LIB3069-004-Q1-K1-G11 LIB3069 g1532072 BLASTN 619 1e−40 90449 -L30783194 LIB3078-052-Q1-K1-E4 LIB3078 g1532072 BLASTN 447 1e−40 76450 -L30792336 LIB3079-021-Q1-K1-E12 LIB3079 g1532072 BLASTN 217 1e−1380 451 1471 700106762H1 SATMON010 g1532072 BLASTN 323 1e−59 84 452 1471701163744H1 SATMONN04 g1532072 BLASTN 220 1e−41 86 453 1471LIB3059-023-Q1-K1-A11 LIB3059 g1532072 BLASTN 340 1e−39 80 454 1471700574530H1 SATMON030 g1532072 BLASTN 247 1e−34 82 455 1471LIB83-003-Q1-E1-G1 LIB83 g1532072 BLASTN 239 1e−24 80 456 1471701161147H1 SATMONN04 g1532072 BLASTN 239 1e−16 80 457 1471 700477336H1SATMON025 g1532072 BLASTN 235 1e−10 91 458 16729 700169502H1 SATMON013g1403043 BLASTN 456 1e−28 65 459 16729 700088201H1 SATMON011 g1532048BLASTX 123 1e−22 60 460 16729 700165440H1 SATMON013 g1532048 BLASTX 1621e−15 65 461 16866 700072905H1 SATMON007 g1532047 BLASTN 368 1e−19 71462 16866 700350558H1 SATMON023 g1532047 BLASTN 301 1e−14 70 463 16866700088370H1 SATMON011 g1532047 BLASTN 306 1e−14 70 464 2324LIB3066-042-Q1-K1-H2 LIB3066 g1403043 BLASTN 1344 1e−103 79 465 2324LIB3059-01 1-Q1-K1-B2 LIB3059 g1403043 BLASTN 1351 1e−103 78 466 2324LIB3059-042-Q1-K1-B7 LIB3059 g1403043 BLASTN 1230 1e−100 80 467 2324LIB3062-036-Q1-K1-F2 LIB3062 g1532072 BLASTN 1097 1e−91 80 468 2324LIB3069-020-Q1-K1-A5 LIB3069 g1403043 BLASTN 925 1e−82 80 469 2324LIB189-023-Q1-E1-H3 LIB189 g1532072 BLASTN 1085 1e−81 78 470 2324700266088H1 SATMON017 g1532047 BLASTN 998 1e−74 80 471 2324LIB3062-026-Q1-K1-E6 LIB3062 g1532047 BLASTN 982 1e−72 77 472 2324700083335H1 SATMON011 g1403043 BLASTN 960 1e−71 79 473 2324LIB189-015-Q1-E1-D3 LIB189 g1532047 BLASTN 970 1e−71 77 474 2324LIB3079-013-Q1-K1-B3 LIB3079 g1403043 BLASTN 953 1e−70 81 475 2324700262129H1 SATMON017 g1403043 BLASTN 936 1e−69 79 476 2324 700265667H1SATMON017 g1403043 BLASTN 694 1e−66 81 477 2324 700807255H1 SATMON036g1532072 BLASTN 850 1e−62 81 478 2324 700196129H1 SATMON014 g1403043BLASTN 853 1e−62 83 479 2324 LIB3066-042-Q1-K1-H1 LIB3066 g1403043BLASTN 842 1e−61 80 480 2324 700458321H1 SATMON029 g1403043 BLASTN 5011e−58 80 481 2324 700197643H1 SATMON014 g1403043 BLASTN 461 1e−57 80 4822324 LIB143-029-Q1-E1-H3 LIB143 g1532072 BLASTN 786 1e−56 78 483 2324700197842H1 SATMON014 g1403043 BLASTN 688 1e−48 85 484 2324 700196807H1SATMON014 g1532072 BLASTN 678 1e−47 75 485 2324 700263712H1 SATMON017g1532072 BLASTN 403 1e−45 75 486 2324 LIB143-006-Q1-E1-F9 LIB143g1403043 BLASTN 324 1e−42 83 487 2324 700267496H1 SATMON017 g1532047BLASTN 594 1e−40 83 488 2324 700172551H1 SATMON013 g1532072 BLASTN 5461e−36 74 489 2324 700211893H1 SATMON016 g1532047 BLASTN 542 1e−35 82 4902324 700264065H1 SATMON017 g1532047 BLASTN 515 1e−34 83 491 2324700465305H1 SATMON025 g1532073 BLASTX 148 1e−26 61 492 2324 700455052H1SATMON029 g1403043 BLASTN 274 1e−26 79 493 2324 700473305H1 SATMON025g1403043 BLASTN 446 1e−26 74 494 2324 700475851H1 SATMON025 g1532047BLASTN 333 1e−18 78 495 2324 700263111H1 SATMON017 g1532073 BLASTX 1651e−15 72 496 2324 700465705H1 SATMON025 g1403044 BLASTX 78 1e−8 76 4973185 700264424H1 SATMON017 g1403043 BLASTN 469 1e−41 81 498 3185LIB143-007-Q1-E1-H2 LIB143 g1403043 BLASTN 464 1e−36 80 499 3185700262548H1 SATMON017 g1532047 BLASTN 368 1e−34 80 500 3185 700263845H1SATMON017 g1403043 BLASTN 455 1e−34 81 501 3185 LIB3068-025-Q1-K1-G10LIB3068 g1403043 BLASTN 288 1e−32 81 502 3185 700264569H1 SATMON017g1403043 BLASTN 469 1e−32 82 503 3185 700243571H1 SATMON010 g1532047BLASTN 461 1e−28 77 504 3185 700265453H1 SATMON017 g1532047 BLASTN 2691e−27 84 505 3185 700267779H1 SATMON017 g1532047 BLASTN 257 1e−26 85 5063185 700382373H1 SATMON024 g1532047 BLASTN 255 1e−24 86 507 3185700258727H1 SATMON017 g1532047 BLASTN 255 1e−24 84 508 3185LIB3069-018-Q1-K1-F5 LIB3069 g1532047 BLASTN 255 1e−21 77 509 3185LIB3066-038-Q1-K1-C2 LIB3066 g1403043 BLASTN 242 1e−17 68 510 3185700334654H1 SATMON019 g1532047 BLASTN 248 1e−17 90 511 3185 700262830H1SATMON017 g1403043 BLASTN 276 1e−12 78 512 3185 700264727H1 SATMON017g1532047 BLASTN 255 1e−10 92 513 3185 700238407H1 SATMON010 g1532047BLASTN 255 1e−10 92 514 3185 700441952H1 SATMON026 g1532047 BLASTN 2551e−10 92 515 3185 700268188H1 SATMON017 g1532047 BLASTN 255 1e−10 92 5163185 700801627H1 SATMON036 g1532047 BLASTN 255 1e−10 92 517 3185700261609H1 SATMON017 g1532047 BLASTN 255 1e−10 92 518 3185 700258510H1SATMON017 g1532047 BLASTN 255 1e−10 92 519 3185 700258779H1 SATMON017g1532047 BLASTN 255 1e−10 92 520 3185 700256838H1 SATMON017 g1532047BLASTN 255 1e−10 92 521 3185 700263361H1 SATMON017 g1532047 BLASTN 2551e−10 92 522 3185 700257102H1 SATMON017 g1532047 BLASTN 255 1e−10 92 5233185 700239452H1 SATMON010 g1532047 BLASTN 245 1e−9 91 524 3185700262045H1 SATMON017 g1532047 BLASTN 250 1e−9 90 525 8LIB3066-027-Q1-K1-C6 LIB3066 g1532072 BLASTN 1414 1e−181 97 526 8LIB3066-048-Q1-K1-C9 LIB3066 g1532072 BLASTN 1715 1e−173 98 527 8LIB3066-007-Q1-K1-B4 LIB3066 g1532072 BLASTN 2146 1e−170 97 528 8LIB3066-019-Q1-K1-B9 LIB3066 g1532072 BLASTN 1884 1e−168 98 529 8LIB148-038-Q1-E1-A3 LIB148 g1532072 BLASTN 1585 1e−164 99 530 8LIB3068-002-Q1-K1-H5 LIB3068 g1532072 BLASTN 2051 1e−162 98 531 8LIB3067-035-Q1-K1-E9 LIB3067 g1532072 BLASTN 1594 1e−161 98 532 8LIB189-020-Q1-E1-E8 LIB189 g1532072 BLASTN 1501 1e−160 98 533 8LIB3078-057-Q1-K1-C12 LIB3078 g1532072 BLASTN 2026 1e−160 96 534 8LIB3066-053-Q1-K1-A8 LIB3066 g1532072 BLASTN 1713 1e−159 93 535 8LIB189-006-Q1-E1-C6 LIB189 g1532072 BLASTN 1676 1e−157 98 536 8LIB3078-054-Q1-K1-E4 LIB3078 g1532072 BLASTN 1997 1e−157 95 537 8LIB3069-028-Q1-K1-A11 LIB3069 g1532072 BLASTN 1980 1e−156 96 538 8LIB3069-045-Q1-K1-H7 LIB3069 g1532072 BLASTN 1989 1e−156 98 539 8LIB148-025-Q1-E1-F7 LIB148 g1532072 BLASTN 1954 1e−154 97 540 8LIB189-014-Q1-E1-F11 LIB189 g1532072 BLASTN 1925 1e−151 94 541 8LIB3060-014-Q1-K1-C5 LIB3060 g1532072 BLASTN 1780 1e−150 96 542 8LIB148-057-Q1-E1-C2 LIB148 g1532072 BLASTN 1000 1e−149 98 543 8LIB148-015-Q1-E1-F2 LIB148 g1532072 BLASTN 1886 1e−148 94 544 8LIB3066-045-Q1-K1-A2 LIB3066 g1532072 BLASTN 914 1e−146 91 545 8LIB148-064-Q1-E1-A3 LIB148 g1532072 BLASTN 1660 1e−146 93 546 8LIB3061-047-Q1-K1-G1 LIB3061 g1532072 BLASTN 1867 1e−146 92 547 8LIB148-040-Q1-E1-F1 LIB148 g1532072 BLASTN 1706 1e−145 94 548 8LIB3069-012-Q1-K1-B6 LIB3069 g1532072 BLASTN 1630 1e−144 87 549 8LIB3068-009-Q1-K1-A8 LIB3068 g1532072 BLASTN 1802 1e−141 97 550 8LIB148-004-Q1-E1-D2 LIB148 g1532072 BLASTN 818 1e−139 91 551 8LIB3068-002-Q1-K1-A1 LIB3068 g1532072 BLASTN 944 1e−137 95 552 8LIB3067-035-Q1-K1-H9 LIB3067 g1532072 BLASTN 1701 1e−137 98 553 8LIB3068-033-Q1-K1-G12 LIB3068 g1532072 BLASTN 1093 1e−130 89 554 8700572229H1 SATMON030 g1532072 BLASTN 1135 1e−128 99 555 8LIB3078-052-Q1-K1-E1 LIB3078 g1532072 BLASTN 1235 1e−127 85 556 8700572579H1 SATMON030 g1532072 BLASTN 1625 1e−126 98 557 8LIB3066-025-Q1-K1-F2 LIB3066 g1532072 BLASTN 1625 1e−126 100 558 8LIB3068-048-Q1-K1-F9 LIB3068 g1532072 BLASTN 1538 1e−125 98 559 8700098413H1 SATMON009 g1532072 BLASTN 1595 1e−124 100 560 8 700573235H1SATMON030 g1532072 BLASTN 1513 1e−123 98 561 8 700090946H1 SATMON011g1532072 BLASTN 1585 1e−123 100 562 8 700092465H1 SATMON008 g1532072BLASTN 1541 1e−122 98 563 8 700074625H1 SATMON007 g1532072 BLASTN 15701e−122 100 564 8 LIB3059-007-Q1-K1-C10 LIB3059 g1532072 BLASTN 14011e−119 94 565 8 700072828H1 SATMON007 g1532072 BLASTN 1540 1e−119 100566 8 700619106H1 SATMON034 g1532072 BLASTN 833 1e−118 97 567 8700074853H1 SATMON007 g1532072 BLASTN 1515 1e−117 100 568 8 700201293H1SATMON003 g1532072 BLASTN 1517 1e−117 97 569 8 700075896H1 SATMON007g1532072 BLASTN 1006 1e−115 99 570 8 LIB3059-052-Q1-K1-A1 LIB3059g1532072 BLASTN 1241 1e−115 92 571 8 700091576H1 SATMON011 g1532072BLASTN 943 1e−114 98 572 8 LIB3078-015-Q1-K1-D7 LIB3078 g1532072 BLASTN1218 1e−114 87 573 8 700074733H1 SATMON007 g1532072 BLASTN 1475 1e−114100 574 8 700381421H1 SATMON023 g1532072 BLASTN 1475 1e−114 100 575 8700085594H1 SATMON011 g1532072 BLASTN 1477 1e−114 99 576 8 700095883H1SATMON008 g1532072 BLASTN 1477 1e−114 99 577 8 700338237H1 SATMON020g1532072 BLASTN 1478 1e−114 99 578 8 700549813H1 SATMON022 g1532072BLASTN 1481 1e−114 99 579 8 700097935H1 SATMON009 g1532072 BLASTN 14841e−114 98 580 8 700572978H1 SATMON030 g1532072 BLASTN 865 1e−113 96 5818 700196464H1 SATMON014 g1532072 BLASTN 1044 1e−113 92 582 8 700381412H1SATMON023 g1532072 BLASTN 1168 1e−113 98 583 8 700027839H1 SATMON003g1532072 BLASTN 1472 1e−113 99 584 8 700623344H1 SATMON034 g1532072BLASTN 1085 1e−112 94 585 8 700025858H1 SATMON003 g1532072 BLASTN 14551e−112 100 586 8 LIB3059-017-Q1-K1-H5 LIB3059 g1532072 BLASTN 14591e−112 97 587 8 700475019H1 SATMON025 g1532072 BLASTN 1390 1e−111 100588 8 700338357H1 SATMON020 g1532072 BLASTN 1440 1e−111 100 589 8700256796H1 SATMON017 g1532072 BLASTN 1400 1e−110 100 590 8 700071692H1SATMON007 g1532072 BLASTN 1430 1e−110 98 591 8 700339073H1 SATMON020g1532072 BLASTN 1357 1e−109 95 592 8 700106916H1 SATMON010 g1532072BLASTN 1415 1e−109 93 593 8 700468234H1 SATMON025 g1532072 BLASTN 14201e−109 100 594 8 700214482H1 SATMON016 g1532072 BLASTN 1425 1e−109 100595 8 700466437H1 SATMON025 g1532072 BLASTN 1403 1e−108 98 596 8700205576H1 SATMON003 g1532072 BLASTN 1404 1e−108 98 597 8 700043455H1SATMON004 g1532072 BLASTN 1405 1e−108 100 598 8 700348439H1 SATMON023g1532072 BLASTN 1407 1e−108 99 599 8 700093131H1 SATMON008 g1532072BLASTN 755 1e−107 100 600 8 700571851H1 SATMON030 g1532072 BLASTN 13021e−107 99 601 8 700088142H1 SATMON011 g1532072 BLASTN 1392 1e−107 99 6028 700085934H1 SATMON011 g1532072 BLASTN 1397 1e−107 98 603 8 700028465H1SATMON003 g1532072 BLASTN 1258 1e−106 98 604 8 700236818H1 SATMON010g1532072 BLASTN 1380 1e−106 100 605 8 700583691H1 SATMON031 g1532072BLASTN 1382 1e−106 99 606 8 700095585H1 SATMON008 g1532072 BLASTN 13831e−106 94 607 8 700105140H1 SATMON010 g1532072 BLASTN 1375 1e−105 100608 8 700338486H1 SATMON020 g1532072 BLASTN 893 1e−104 98 609 8700214178H1 SATMON016 g1532072 BLASTN 1361 1e−104 99 610 8 700090613H1SATMON011 g1532072 BLASTN 699 1e−103 99 611 8 700576189H1 SATMON030g1532072 BLASTN 1254 1e−103 93 612 8 700475734H1 SATMON025 g1532072BLASTN 1256 1e−103 99 613 8 700028218H1 SATMON003 g1532072 BLASTN 12751e−103 100 614 8 700088644H1 SATMON011 g1532072 BLASTN 1351 1e−103 98615 8 700378768H1 SATMON020 g1532072 BLASTN 1067 1e−102 98 616 8LIB3067-035-Q1-K1-H10 LIB3067 g1532072 BLASTN 1233 1e−102 95 617 8700043466H1 SATMON004 g1532072 BLASTN 1331 1e−102 97 618 8LIB148-057-Q1-E1-C3 LIB148 g1532072 BLASTN 1283 1e−101 92 619 8700217574H1 SATMON016 g1532072 BLASTN 1320 1e−101 100 620 8 700042332H1SATMON004 g1532072 BLASTN 1321 1e−101 97 621 8 700440926H1 SATMON026g1532072 BLASTN 1323 1e−101 99 622 8 700552284H1 SATMON022 g1532072BLASTN 1329 1e−101 97 623 8 700216613H1 SATMON016 g1532072 BLASTN 6891e−100 97 624 8 LIB3067-054-Q1-K1-F6 LIB3067 g1532072 BLASTN 1030 1e−10094 625 8 LIB3060-038-Q1-K1-B9 LIB3060 g1532072 BLASTN 1108 1e−100 92 6268 700218755H1 SATMON011 g1532072 BLASTN 1116 1e−100 99 627 8 700338042H1SATMON020 g1532072 BLASTN 1270 1e−100 99 628 8 700268009H1 SATMON017g1532072 BLASTN 1307 1e−100 92 629 8 700578491H1 SATMON031 g1532072BLASTN 1309 1e−100 97 630 8 700030037H1 SATMON003 g1532072 BLASTN 13101e−100 97 631 8 700221406H1 SATMON011 g1532072 BLASTN 1315 1e−100 100632 8 700157357H1 SATMON012 g1532072 BLASTN 1315 1e−100 98 633 8700476926H1 SATMON025 g1532072 BLASTN 680 1e−99 97 634 8 700475068H1SATMON025 g1532072 BLASTN 792 1e−99 98 635 8 700469741H1 SATMON025g1532072 BLASTN 831 1e−99 99 636 8 700082377H1 SATMON011 g1532072 BLASTN1081 1e−99 99 637 8 700217143H1 SATMON016 g1532072 BLASTN 1208 1e−99 98638 8 700339433H1 SATMON020 g1532072 BLASTN 1295 1e−99 100 639 8700196627H1 SATMON014 g1532072 BLASTN 1295 1e−99 100 640 8 700259354H1SATMON017 g1532072 BLASTN 1305 1e−99 92 641 8 700610842H1 SATMON022g1532072 BLASTN 843 1e−98 97 642 8 700218059H1 SATMON016 g1532072 BLASTN1036 1e−98 99 643 8 700421727H1 SATMONN01 g1532072 BLASTN 1172 1e−98 97644 8 700158660H1 SATMON012 g1532072 BLASTN 1285 1e−98 100 645 8700806856H1 SATMON036 g1532072 BLASTN 1194 1e−97 97 646 8 700583512H1SATMON031 g1532072 BLASTN 1219 1e−97 97 647 8 700025502H1 SATMON004g1532072 BLASTN 1276 1e−97 99 648 8 700159562H1 SATMON012 g1532072BLASTN 1277 1e−97 99 649 8 700223731H1 SATMON011 g1532072 BLASTN 12771e−97 99 650 8 700156494H1 SATMON012 g1532072 BLASTN 1280 1e−97 100 6518 700160533H1 SATMON012 g1532072 BLASTN 1281 1e−97 97 652 8LIB3066-055-Q1-K1-D8 LIB3066 g1532072 BLASTN 672 1e−96 99 653 8700405152H1 SATMON028 g1532072 BLASTN 828 1e−96 98 654 8LIB148-040-Q1-E1-A8 LIB148 g1532072 BLASTN 1268 1e−96 83 655 8700438437H1 SATMON026 g1532072 BLASTN 1246 1e−95 99 656 8 700267245H1SATMON017 g1532072 BLASTN 1248 1e−95 93 657 8 700156129H2 SATMON007g1532072 BLASTN 1250 1e−95 100 658 8 700203246H1 SATMON003 g1532072BLASTN 1250 1e−95 100 659 8 700168339H1 SATMON013 g1532072 BLASTN 9151e−94 98 660 8 LIB3067-036-Q1-K1-F9 LIB3067 g1532072 BLASTN 1110 1e−9495 661 8 700193769H1 SATMON014 g1532072 BLASTN 1240 1e−94 100 662 8700094014H1 SATMON008 g1532072 BLASTN 1241 1e−94 91 663 8 700020311H1SATMON001 g1532072 BLASTN 1228 1e−93 99 664 8 700195083H1 SATMON014g1532072 BLASTN 1229 1e−93 98 665 8 700193901H1 SATMON014 g1532072BLASTN 1229 1e−93 98 666 8 700194639H1 SATMON014 g1532072 BLASTN 12311e−93 99 667 8 700350117H1 SATMON023 g1532072 BLASTN 1143 1e−92 99 668 8700239867H1 SATMON010 g1532072 BLASTN 1210 1e−92 98 669 8LIB3069-028-Q1-K1-D1 LIB3069 g1532072 BLASTN 1217 1e−92 94 670 8700355756H1 SATMON024 g1532072 BLASTN 618 1e−91 96 671 8 700265492H1SATMON017 g1532072 BLASTN 620 1e−91 94 672 8 700579021H1 SATMON031g1532072 BLASTN 656 1e−91 96 673 8 LIB3079-015-Q1-K1-B11 LIB3079g1532072 BLASTN 1008 1e−91 83 674 8 700572888H2 SATMON030 g1532072BLASTN 1159 1e−91 99 675 8 701183990H1 SATMONN06 g1532072 BLASTN 11981e−91 93 676 8 700085757H1 SATMON011 g1532072 BLASTN 1200 1e−91 100 6778 700162756H1 SATMON013 g1532072 BLASTN 1208 1e−91 99 678 8 700193076H1SATMON014 g1532072 BLASTN 1208 1e−91 99 679 8 700224378H1 SATMON011g1532072 BLASTN 696 1e−90 99 680 8 700218773H1 SATMON011 g1532072 BLASTN1187 1e−90 93 681 8 700159338H1 SATMON012 g1532072 BLASTN 1193 1e−90 97682 8 700169217H1 SATMON013 g1532072 BLASTN 1196 1e−90 99 683 8700551548H1 SATMON022 g1532072 BLASTN 839 1e−89 96 684 8 700197059H1SATMON014 g1532072 BLASTN 1025 1e−89 95 685 8 700469834H1 SATMON025g1532072 BLASTN 650 1e−88 97 686 8 700569778H1 SATMON030 g1532072 BLASTN682 1e−88 91 687 8 700194845H1 SATMON014 g1532072 BLASTN 1171 1e−88 99688 8 700445861H1 SATMON027 g1532072 BLASTN 536 1e−87 99 689 8700100536H1 SATMON009 g1532072 BLASTN 666 1e−87 94 690 8 701163801H1SATMONN04 g1532072 BLASTN 805 1e−87 94 691 8 LIB3060-035-Q1-K1-E3LIB3060 g1532072 BLASTN 922 1e−87 95 692 8 700170872H1 SATMON013g1532072 BLASTN 943 1e−87 97 693 8 700241270H1 SATMON010 g1532072 BLASTN1066 1e−86 96 694 8 700457981H1 SATMON029 g1532072 BLASTN 1140 1e−86 92695 8 700158339H1 SATMON012 g1532072 BLASTN 1142 1e−86 99 696 8700019442H1 SATMON001 g1532072 BLASTN 1145 1e−86 100 697 8 700149885H1SATMON007 g1532072 BLASTN 1133 1e−85 99 698 8 700018255H1 SATMON001g1532072 BLASTN 1135 1e−85 100 699 8 700244026H1 SATMON010 g1532072BLASTN 1044 1e−84 89 700 8 LIB3060-027-Q1-K1-B2 LIB3060 g1532072 BLASTN1117 1e−84 99 701 8 700170960H1 SATMON013 g1532072 BLASTN 696 1e−83 99702 8 700267487H1 SATMON017 g1532072 BLASTN 1061 1e−83 92 703 8700152754H1 SATMON007 g1532072 BLASTN 1107 1e−83 99 704 8 700378260H1SATMON019 g1532072 BLASTN 1112 1e−83 92 705 8 700455089H1 SATMON029g1532072 BLASTN 583 1e−82 97 706 8 700167322H1 SATMON013 g1532072 BLASTN1080 1e−81 98 707 8 LIB3060-035-Q1-K1-H1 LIB3060 g1532072 BLASTN 7821e−80 94 708 8 700204067H1 SATMON003 g1532072 BLASTN 1011 1e−80 98 709 8700049271H1 SATMON003 g1532072 BLASTN 627 1e−79 93 710 8 700442065H1SATMON026 g1532072 BLASTN 1043 1e−78 91 711 8 700244175H1 SATMON010g1532072 BLASTN 1044 1e−78 90 712 8 700045296H1 SATMON004 g1532072BLASTN 1041 1e−77 93 713 8 700578391H1 SATMON031 g1532072 BLASTN 6241e−76 88 714 8 700346136H1 SATMON021 g1532072 BLASTN 751 1e−76 90 715 8700457852H1 SATMON029 g1532072 BLASTN 777 1e−76 92 716 8 700570111H1SATMON030 g1532072 BLASTN 814 1e−75 94 717 8 LIB3066-030-Q1-K1-A12LIB3066 g1532072 BLASTN 922 1e−75 94 718 8 700149657H1 SATMON007g1532072 BLASTN 1012 1e−75 92 719 8 LIB3060-043-Q1-K1-B4 LIB3060g1532072 BLASTN 1013 1e−75 97 720 8 700156752H1 SATMON012 g1532072BLASTN 1013 1e−75 98 721 8 700454114H1 SATMON029 g1532072 BLASTN 4551e−74 93 722 8 LIB143-053-Q1-E1-E8 LIB143 g1532072 BLASTN 672 1e−74 98723 8 700029323H1 SATMON003 g1532072 BLASTN 996 1e−74 99 724 8700156381H1 SATMON007 g1532072 BLASTN 1002 1e−74 97 725 8 700159603H2SATMON012 g1532072 BLASTN 1003 1e−74 94 726 8 LIB3060-035-Q1-K1-E5LIB3060 g1532072 BLASTN 603 1e−72 94 727 8 LIB3069-025-Q1-K1-E12 LIB3069g1532072 BLASTN 734 1e−72 90 728 8 700166680H1 SATMON013 g1532072 BLASTN971 1e−72 99 729 8 700171123H1 SATMON013 g1532072 BLASTN 973 1e−72 94730 8 LIB148-042-Q1-E1-G10 LIB148 g1532072 BLASTN 975 1e−72 100 731 8700612951H1 SATMON033 g1532072 BLASTN 401 1e−71 96 732 8 700265213H1SATMON017 g1532072 BLASTN 626 1e−71 92 733 8 LIB189-030-Q1-E1-D11 LIB189g1532072 BLASTN 740 1e−70 92 734 8 700212877H1 SATMON016 g1532072 BLASTN918 1e−70 93 735 8 700161545H1 SATMON012 g1532072 BLASTN 621 1e−69 99736 8 700021259H1 SATMON001 g1532072 BLASTN 940 1e−69 93 737 8LIB3079-015-Q1-K1-C7 LIB3079 g1532072 BLASTN 508 1e−68 96 738 8LIB3066-055-Q1-K1-F11 LIB3066 g1532072 BLASTN 832 1e−68 85 739 8700166771H1 SATMON013 g1532072 BLASTN 922 1e−68 88 740 8LIB3066-003-Q1-K1-E6 LIB3066 g1532072 BLASTN 927 1e−68 86 741 8700208131H1 SATMON016 g1532047 BLASTN 747 1e−66 85 742 8 700020688H1SATMON001 g1532072 BLASTN 905 1e−66 90 743 8 700206405H1 SATMON003g1532072 BLASTN 905 1e−66 100 744 8 LIB3069-042-Q1-K1-C5 LIB3069g1532072 BLASTN 891 1e−65 99 745 8 700353835H1 SATMON024 g1532072 BLASTN896 1e−65 96 746 8 700471533H1 SATMON025 g1532072 BLASTN 496 1e−63 99747 8 700165425H1 SATMON013 g1532072 BLASTN 850 1e−62 100 748 8LIB3069-054-Q1-K1-C4 LIB3069 g1532072 BLASTN 613 1e−60 87 749 8700160896H1 SATMON012 g1532072 BLASTN 746 1e−60 95 750 8LIB3069-042-Q1-K1-A11 LIB3069 g1532072 BLASTN 840 1e−60 98 751 8700466625H1 SATMON025 g1532072 BLASTN 635 1e−59 86 752 8 700571770H1SATMON030 g1532072 BLASTN 820 1e−59 84 753 8 LIB143-024-Q1-E1-D2 LIB143g1532072 BLASTN 813 1e−58 95 754 8 700453677H1 SATMON028 g1532072 BLASTN508 1e−57 92 755 8 700193979H1 SATMON014 g1532072 BLASTN 790 1e−57 100756 8 700467828H1 SATMON025 g1532072 BLASTN 779 1e−56 96 757 8700150072H1 SATMON007 g1532072 BLASTN 776 1e−55 99 758 8 700802869H1SATMON036 g1532072 BLASTN 702 1e−54 95 759 8 LIB148-051-Q1-E1-B12 LIB148g1532072 BLASTN 421 1e−53 92 760 8 700075035H1 SATMON007 g1532072 BLASTN506 1e−53 93 761 8 700471283H1 SATMON025 g1532072 BLASTN 742 1e−53 91762 8 700166625H1 SATMON013 g1532072 BLASTN 748 1e−53 99 763 8700019894H1 SATMON001 g1532072 BLASTN 643 1e−51 89 764 8 700204863H1SATMON003 g1532072 BLASTN 721 1e−51 99 765 8 700431071H1 SATMONN01g1532072 BLASTN 366 1e−50 94 766 8 700171582H1 SATMON013 g1532072 BLASTN492 1e−50 99 767 8 700450579H1 SATMON028 g1532072 BLASTN 338 1e−49 97768 8 700084149H1 SATMON011 g1532072 BLASTN 672 1e−47 98 769 8700095070H1 SATMON008 g1532072 BLASTN 662 1e−46 98 770 8 700570827H1SATMON030 g1532072 BLASTN 344 1e−44 83 771 8 LIB3061-057-Q1-K1-G12LIB3061 g1532072 BLASTN 378 1e−43 75 772 8 700194918H1 SATMON014g1532072 BLASTN 626 1e−43 89 773 8 700378894H1 SATMON020 g1532072 BLASTN495 1e−42 97 774 8 700468029H1 SATMON025 g1532072 BLASTN 616 1e−42 98775 8 LIB143-012-Q1-E1-H8 LIB143 g1532072 BLASTN 637 1e−42 98 776 8LIB3060-037-Q1-K1-H4 LIB3060 g1532072 BLASTN 638 1e−42 86 777 8700433327H1 SATMONN01 g1532072 BLASTN 365 1e−41 86 778 8 700623611H1SATMON034 g1532072 BLASTN 316 1e−39 95 779 8 700166123H1 SATMON013g1532072 BLASTN 585 1e−39 100 780 8 700616273H1 SATMON033 g1532072BLASTN 534 1e−38 98 781 8 700464702H1 SATMON025 g1532072 BLASTN 5661e−38 99 782 8 700338809H1 SATMON020 g1532072 BLASTN 550 1e−37 100 783 8700092622H1 SATMON008 g1532072 BLASTN 561 1e−37 99 784 8LIB3060-043-Q1-K1-A10 LIB3060 g1532072 BLASTN 352 1e−36 93 785 8700265611H1 SATMON017 g1532072 BLASTN 548 1e−36 95 786 8 700100319H1SATMON009 g1532072 BLASTN 552 1e−36 98 787 8 700092696H1 SATMON008g1532072 BLASTN 552 1e−36 98 788 8 700076750H1 SATMON007 g1532072 BLASTN526 1e−35 99 789 8 700082896H1 SATMON011 g1532072 BLASTN 526 1e−35 99790 8 700075411H1 SATMON007 g1532072 BLASTN 319 1e−34 91 791 8701178051H1 SATMONN05 g1532072 BLASTN 342 1e−34 94 792 8 700266358H1SATMON017 g1532072 BLASTN 520 1e−34 95 793 8 700453981H1 SATMON029g1532072 BLASTN 505 1e−33 96 794 8 700584238H1 SATMON031 g1532072 BLASTN509 1e−33 91 795 8 700103871H1 SATMON010 g1532072 BLASTN 491 1e−32 99796 8 700264460H1 SATMON017 g1532072 BLASTN 495 1e−32 95 797 8700205122H1 SATMON003 g1532072 BLASTN 501 1e−32 99 798 8 700165768H1SATMON013 g1532072 BLASTN 420 1e−31 98 799 8 700077179H1 SATMON007g1532072 BLASTN 471 1e−30 98 800 8 700476654H1 SATMON025 g1532072 BLASTN476 1e−30 98 801 8 700027374H1 SATMON003 g1532072 BLASTN 476 1e−30 98802 8 700266994H1 SATMON017 g1532072 BLASTN 476 1e−30 98 803 8700088193H1 SATMON011 g1532072 BLASTN 488 1e−30 97 804 8 700214511H1SATMON016 g1532072 BLASTN 298 1e−29 93 805 8 700257186H1 SATMON017g1532072 BLASTN 465 1e−29 95 806 8 700335814H1 SATMON019 g1532072 BLASTN469 1e−29 93 807 8 700236166H1 SATMON010 g1532072 BLASTN 450 1e−28 95808 8 700468243H1 SATMON025 g1532072 BLASTN 456 1e−28 98 809 8700096327H1 SATMON008 g1532072 BLASTN 451 1e−27 98 810 8 700471139H1SATMON025 g1532072 BLASTN 451 1e−27 98 811 8 700050420H1 SATMON003g1532072 BLASTN 365 1e−26 92 812 8 700264846H1 SATMON017 g1532072 BLASTN430 1e−25 94 813 8 700266803H1 SATMON017 g1532072 BLASTN 293 1e−24 75814 8 700086134H1 SATMON011 g1532072 BLASTN 422 1e−24 97 815 8700162001H1 SATMON012 g1532072 BLASTN 279 1e−23 96 816 8 700044825H1SATMON004 g1532072 BLASTN 411 1e−23 98 817 8 700074679H1 SATMON007g1532072 BLASTN 411 1e−23 98 818 8 700267694H1 SATMON017 g1532072 BLASTN320 1e−22 92 819 8 700267990H1 SATMON017 g1532072 BLASTN 361 1e−22 99820 8 700456715H1 SATMON029 g1532072 BLASTN 293 1e−21 97 821 8700454806H1 SATMON029 g1532072 BLASTN 356 1e−21 98 822 8 700466812H1SATMON025 g1532072 BLASTN 380 1e−21 95 823 8 700046476H1 SATMON004g1532072 BLASTN 381 1e−21 97 824 8 700207160H1 SATMON017 g1532072 BLASTN393 1e−21 96 825 8 700383037H1 SATMON024 g1532072 BLASTN 280 1e−20 98826 8 700549660H1 SATMON022 g1532072 BLASTN 376 1e−20 97 827 8LIB3066-004-Q1-K1-G12 LIB3066 g1532072 BLASTN 379 1e−20 91 828 8700801422H1 SATMON036 g1532072 BLASTN 321 1e−19 97 829 8 700045319H1SATMON004 g1532072 BLASTN 336 1e−17 99 830 8 700046047H1 SATMON004g1532072 BLASTN 336 1e−17 99 831 8 700264328H1 SATMON017 g1532072 BLASTN339 1e−17 95 832 8 700083485H1 SATMON011 g1532072 BLASTN 341 1e−17 99833 8 700461268H1 SATMON033 g1532072 BLASTN 342 1e−17 95 834 8700053271H1 SATMON008 g1532072 BLASTN 311 1e−15 98 835 8 700215275H1SATMON016 g1532072 BLASTN 321 1e−15 98 836 8 700623133H1 SATMON034g1532072 BLASTN 275 1e−14 75 837 8 700454949H1 SATMON029 g1532072 BLASTN280 1e−14 99 838 8 700440763H1 SATMON026 g1532072 BLASTN 307 1e−14 95839 8 LIB3068-022-Q1-K1-C4 LIB3068 g1532072 BLASTN 150 1e−12 94 840 8700798849H1 SATMON036 g1532072 BLASTN 281 1e−12 98 841 8 700333057H1SATMON019 g1532072 BLASTN 184 1e−11 91 842 8 700265786H1 SATMON017g1532072 BLASTN 200 1e−11 90 843 8 700193030H1 SATMON014 g1532072 BLASTN158 1e−9 94 844 8 700262964H1 SATMON017 g1532047 BLASTN 171 1e−9 81 8458 LIB3069-023-Q1-K1-B2 LIB3069 g1403043 BLASTN 195 1e−9 84 846 8700474096H1 SATMON025 g1532072 BLASTN 199 1e−9 97 847 8 700028326H1SATMON003 g1532072 BLASTN 151 1e−8 96 848 8011 700440327H1 SATMON026g1532047 BLASTN 292 1e−14 85 849 851 LIB3059-052-Q1-K1-E6 LIB3059g1403043 BLASTN 1033 1e−85 75 850 851 700090958H1 SATMON011 g1532072BLASTN 1044 1e−78 80 851 851 700224605H1 SATMON011 g1532072 BLASTN 8501e−62 80 852 851 700551562H1 SATMON022 g1403043 BLASTN 374 1e−55 79 853851 700153939H1 SATMON007 g1403043 BLASTN 772 1e−55 80 854 851700469231H1 SATMON025 g1532072 BLASTN 560 1e−52 76 855 851 700349854H1SATMON023 g1403043 BLASTN 669 1e−46 81 856 851 701169324H1 SATMONN05g1403043 BLASTN 669 1e−46 78 857 851 700088319H1 SATMON011 g1532072BLASTN 492 1e−30 78 2480 -700998660 700998660H1 SOYMON018 g1531764BLASTN 249 1e−29 90 2481 -GM927 LIB3028-005-Q1-B1-B4 LIB3028 g1421750BLASTN 261 1e−10 80 2482 13379 700842514H1 SOYMON020 g1915980 BLASTN 5751e−41 73 2483 13379 701042786H1 SOYMON029 g1421750 BLASTN 377 1e−20 802484 15556 701101584H1 SOYMON028 g1421750 BLASTN 526 1e−35 69 2485 15556701099749H1 SOYMON028 g1155239 BLASTN 453 1e−27 72 2486 16LIB3051-018-Q1-E1-A7 LIB3051 g1421750 BLASTN 1146 1e−93 81 2487 16LIB3056-005-Q1-N1-E10 LIB3056 g1421750 BLASTN 944 1e−88 80 2488 16LIB3051-101-Q1-K1-C7 LIB3051 g1421750 BLASTN 1135 1e−85 80 2489 16LIB3056-001-Q1-B1-H12 LIB3056 g1421750 BLASTN 752 1e−82 81 2490 16700661334H1 SOYMON005 g1421750 BLASTN 1092 1e−82 80 2491 16LIB3056-012-Q1-N1-B12 LIB3056 g1421750 BLASTN 1085 1e−81 80 2492 16LIB3055-011-Q1-N1-G2 LIB3055 g1421750 BLASTN 958 1e−78 79 2493 16700663981H1 SOYMON005 g1421750 BLASTN 996 1e−74 82 2494 16 701109737H1SOYMON036 g1421750 BLASTN 955 1e−70 83 2495 16 701130190H1 SOYMON037g1421750 BLASTN 935 1e−69 81 2496 16 701003301H1 SOYMON019 g1421750BLASTN 412 1e−68 82 2497 16 700894137H1 SOYMON024 g1421750 BLASTN 9231e−68 84 2498 16 700980096H1 SOYMON009 g1421750 BLASTN 923 1e−68 81 249916 700942625H1 SOYMON024 g1421750 BLASTN 911 1e−67 80 2500 16700829695H1 SOYMON019 g1421750 BLASTN 918 1e−67 84 2501 16 700662572H1SOYMON005 g1421750 BLASTN 920 1e−67 84 2502 16 701127647H1 SOYMON037g1421750 BLASTN 902 1e−66 84 2503 16 701014550H1 SOYMON019 g1421750BLASTN 556 1e−65 81 2504 16 LIB3051-043-Q1-K1-G3 LIB3051 g1421750 BLASTN698 1e−65 82 2505 16 700730914H1 SOYMON009 g1421750 BLASTN 897 1e−65 842506 16 701052455H1 SOYMON032 g1421750 BLASTN 876 1e−64 81 2507 16LIB3051-074-Q1-K1-A7 LIB3051 g1421750 BLASTN 876 1e−64 75 2508 16700874839H1 SOYMON018 g1421750 BLASTN 862 1e−63 82 2509 16 701005943H1SOYMON019 g1421750 BLASTN 865 1e−63 82 2510 16 700663357H1 SOYMON005g1421750 BLASTN 869 1e−63 82 2511 16 701042053H1 SOYMON029 g1421750BLASTN 872 1e−63 81 2512 16 700971988H1 SOYMON005 g1421750 BLASTN 5081e−61 82 2513 16 701010728H1 SOYMON019 g1421750 BLASTN 657 1e−61 81 251416 700987206H1 SOYMON009 g1421750 BLASTN 845 1e−61 83 2515 16701126773H1 SOYMON037 g1421750 BLASTN 847 1e−61 79 2516 16 700764714H1SOYMON023 g1421750 BLASTN 830 1e−60 81 2517 16 700974444H1 SOYMON005g1421750 BLASTN 830 1e−60 81 2518 16 701118620H1 SOYMON037 g1421750BLASTN 832 1e−60 75 2519 16 700729926H1 SOYMON009 g1421750 BLASTN 8341e−60 79 2520 16 700945142H1 SOYMON024 g1421750 BLASTN 470 1e−59 84 252116 700867912H1 SOYMON016 g1421750 BLASTN 820 1e−59 82 2522 16700746546H1 SOYMON013 g1421750 BLASTN 824 1e−59 78 2523 16 700873331H1SOYMON018 g1421750 BLASTN 769 1e−58 82 2524 16 700738212H1 SOYMON012g1421750 BLASTN 780 1e−56 83 2525 16 700830454H1 SOYMON019 g1421750BLASTN 789 1e−56 81 2526 16 700726129H1 SOYMON009 g1421750 BLASTN 7891e−56 81 2527 16 700996774H1 SOYMON018 g1421750 BLASTN 486 1e−55 81 252816 700746427H1 SOYMON013 g1421750 BLASTN 766 1e−55 77 2529 16701123911H1 SOYMON037 g1421750 BLASTN 769 1e−55 76 2530 16 700745730H1SOYMON013 g1421750 BLASTN 775 1e−55 76 2531 16 700900940H1 SOYMON027g1421750 BLASTN 760 1e−54 78 2532 16 700846419H1 SOYMON021 g1421750BLASTN 762 1e−54 78 2533 16 701203427H1 SOYMON035 g1421750 BLASTN 4911e−53 84 2534 16 LIB3051-074-Q1-K1-F5 LIB3051 g1421750 BLASTN 650 1e−5375 2535 16 700747646H1 SOYMON013 g1421750 BLASTN 747 1e−53 77 2536 16701049823H1 SOYMON032 g1421750 BLASTN 751 1e−53 77 2537 16 701049188H1SOYMON032 g1421750 BLASTN 731 1e−52 75 2538 16 700875156H1 SOYMON018g1421750 BLASTN 737 1e−52 78 2539 16 700864540H1 SOYMON016 g1421750BLASTN 741 1e−52 81 2540 16 700682329H2 SOYMON008 g1421750 BLASTN 7191e−51 76 2541 16 700846215H1 SOYMON021 g1421750 BLASTN 575 1e−50 82 254216 701101327H1 SOYMON028 g1421750 BLASTN 592 1e−50 77 2543 16LIB3055-011-Q1-N1-E7 LIB3055 g1421750 BLASTN 680 1e−50 80 2544 16700848992H1 SOYMON021 g1421750 BLASTN 708 1e−50 76 2545 16 700966820H1SOYMON028 g1421750 BLASTN 710 1e−50 75 2546 16 701051785H1 SOYMON032g1421750 BLASTN 717 1e−50 75 2547 16 700681079H1 SOYMON008 g1917012BLASTN 290 1e−49 77 2548 16 701138880H1 SOYMON038 g1421750 BLASTN 6101e−49 78 2549 16 700872967H1 SOYMON018 g1421750 BLASTN 704 1e−49 75 255016 701119960H1 SOYMON037 g1421750 BLASTN 684 1e−48 81 2551 16700873306H1 SOYMON018 g1421750 BLASTN 693 1e−48 83 2552 16 700751484H1SOYMON014 g1421750 BLASTN 635 1e−47 82 2553 16 700958865H1 SOYMON022g1421750 BLASTN 486 1e−46 82 2554 16 700872833H1 SOYMON018 g1421750BLASTN 659 1e−46 84 2555 16 700871673H1 SOYMON018 g1421750 BLASTN 6591e−46 84 2556 16 700872801H1 SOYMON018 g1421750 BLASTN 659 1e−46 84 255716 700847572H1 SOYMON021 g1421750 BLASTN 660 1e−46 75 2558 16700728046H1 SOYMON009 g1421750 BLASTN 661 1e−46 84 2559 16 700894721H1SOYMON024 g1421750 BLASTN 550 1e−43 84 2560 16 700730946H1 SOYMON009g1421750 BLASTN 307 1e−42 77 2561 16 700727943H1 SOYMON009 g1421750BLASTN 378 1e−42 79 2562 16 701101238H1 SOYMON028 g1421750 BLASTN 4561e−42 92 2563 16 700758519H1 SOYMON015 g1421750 BLASTN 358 1e−41 77 256416 700740343H1 SOYMON012 g1531764 BLASTN 607 1e−41 72 2565 16700848076H1 SOYMON021 g1531764 BLASTN 469 1e−40 89 2566 16LIB3051-080-Q1-K1-H7 LIB3051 g1531764 BLASTN 472 1e−38 90 2567 16700955916H1 SOYMON022 g1531764 BLASTN 427 1e−37 90 2568 16 700873876H1SOYMON018 g1421750 BLASTN 465 1e−37 80 2569 16 700746271H1 SOYMON013g1421750 BLASTN 515 1e−37 80 2570 16 700662887H1 SOYMON005 g1421750BLASTN 506 1e−35 82 2571 16 700983591H1 SOYMON009 g1421750 BLASTN 5371e−35 82 2572 16 700752343H1 SOYMON014 g1421750 BLASTN 488 1e−34 73 257316 700901973H1 SOYMON027 g1421750 BLASTN 490 1e−34 82 2574 16LIB3053-013-Q1-N1-B11 LIB3053 g1421750 BLASTN 522 1e−34 75 2575 16700988481H1 SOYMON009 g1421750 BLASTN 506 1e−33 78 2576 16LIB3051-078-Q1-K1-C12 LIB3051 g1421750 BLASTN 302 1e−32 75 2577 16700985640H1 SOYMON009 g1421750 BLASTN 449 1e−30 86 2578 16 701127038H1SOYMON037 g1421750 BLASTN 467 1e−30 67 2579 16 700898677H1 SOYMON027g1531764 BLASTN 474 1e−30 89 2580 16 700742260H1 SOYMON012 g1421750BLASTN 476 1e−30 70 2581 16 701103203H1 SOYMON028 g1421750 BLASTN 2871e−27 81 2582 16 700897144H1 SOYMON027 g1421750 BLASTN 264 1e−26 75 258316 700832039H1 SOYMON019 g1421750 BLASTN 390 1e−26 82 2584 16700743311H1 SOYMON012 g1421750 BLASTN 429 1e−26 87 2585 16LIB3040-044-Q1-E1-B8 LIB3040 g1421750 BLASTN 282 1e−25 92 2586 16LIB3051-024-Q1-K1-C4 LIB3051 g2394382 BLASTX 110 1e−24 95 2587 16701011765H1 SOYMON019 g1421750 BLASTN 383 1e−21 76 2588 16 700893428H1SOYMON024 g1421752 BLASTX 106 1e−20 82 2589 16 700996204H1 SOYMON018g1421750 BLASTN 288 1e−20 75 2590 16 LIB3049-028-Q1-E1-C6 LIB3049g1421750 BLASTN 230 1e−19 90 2591 16 701103628H1 SOYMON028 g1421750BLASTN 277 1e−18 73 2592 16 700972636H1 SOYMON005 g1421750 BLASTN 2861e−16 73 2593 16 700875417H1 SOYMON018 g1421750 BLASTN 209 1e−15 75 259416 701207702H1 SOYMON035 g1421752 BLASTX 153 1e−14 83 2595 16701054556H1 SOYMON032 g1421750 BLASTN 230 1e−14 75 2596 16 701046407H1SOYMON032 g1421750 BLASTN 230 1e−14 75 2597 16 701117634H1 SOYMON037g1421750 BLASTN 230 1e−14 74 2598 16 701127714H1 SOYMON037 g1421750BLASTN 230 1e−14 75 2599 16 700663239H1 SOYMON005 g1421750 BLASTN 2101e−13 74 2600 16 700738510H1 SOYMON012 g1421750 BLASTN 212 1e−13 72 260116 700943559H1 SOYMON024 g1421750 BLASTN 215 1e−13 71 2602 16700666278H1 SOYMON005 g1421750 BLASTN 290 1e−13 93 2603 16 700663144H1SOYMON005 g1421750 BLASTN 204 1e−12 74 2604 16 700684147H1 SOYMON008g1421750 BLASTN 278 1e−12 89 2605 16 700953076H1 SOYMON022 g1421750BLASTN 285 1e−12 92 2606 16 701099931H1 SOYMON028 g1421750 BLASTN 2021e−11 72 2607 16 701058910H1 SOYMON033 g1421750 BLASTN 216 1e−11 74 260816 700754949H1 SOYMON014 g1421750 BLASTN 266 1e−11 89 2609 16700562967H1 SOYMON002 g1421750 BLASTN 266 1e−11 82 2610 16 701012790H1SOYMON019 g1421750 BLASTN 251 1e−10 89 2611 16 701101739H1 SOYMON028g1421750 BLASTN 260 1e−10 82 2612 16 701118211H1 SOYMON037 g1490554BLASTX 97 1e−9 47 2613 16 700844394H1 SOYMON021 g1917013 BLASTX 116 1e−985 2614 16 701048560H1 SOYMON032 g1421750 BLASTN 198 1e−9 79 2615 16701062373H1 SOYMON033 g1421750 BLASTN 203 1e−9 71 2616 16 701120496H1SOYMON037 g1421750 BLASTN 230 1e−9 82 2617 16 701049680H1 SOYMON032g1421750 BLASTN 177 1e−8 70 2618 16 700748760H1 SOYMON013 g1421750BLASTN 230 1e−8 91 2619 16 700889482H1 SOYMON024 g1421750 BLASTN 2351e−8 91 2620 16 700725013H1 SOYMON009 g1421750 BLASTN 239 1e−8 85 262116048 LIB3028-004-Q1-B1-E2 LIB3028 g1421750 BLASTN 879 1e−64 68 262216048 700761667H1 SOYMON015 g1421750 BLASTN 528 1e−35 71 2623 16048700958003H1 SOYMON022 g1421750 BLASTN 428 1e−26 78

ASPARTATE KINASE (EC 2.7.2.4) Seq No. Cluster ID CloneID Library NCBI giMethod Score P-value % Ident 858 -700018870 700018870H1 SATMON001g500850 BLASTN 1095 1e−82 100 859 -700085903 700085903H1 SATMON011g500852 BLASTN 1606 1e−124 99 860 -700086169 700086169H1 SATMON011g500852 BLASTN 559 1e−94 92 861 -700096679 700096679H1 SATMON008 g500850BLASTN 1131 1e−85 99 862 -700096794 700096794H1 SATMON008 g500850 BLASTN1515 1e−117 100 863 -700106390 700106390H1 SATMON010 g2243115 BLASTN 5441e−51 72 864 -700168286 700168286H1 SATMON013 g2243115 BLASTN 519 1e−3472 865 -700171363 700171363H1 SATMON013 g500850 BLASTN 1085 1e−81 94 866-700194781 700194781H1 SATMON014 g2257742 BLASTN 635 1e−44 79 867-700213839 700213839H1 SATMON016 g500850 BLASTN 664 1e−75 94 868-700219756 700219756H1 SATMON011 g500850 BLASTN 1359 1e−104 99 869-700258808 700258808H1 SATMON017 g500850 BLASTN 630 1e−85 96 870-700263439 700263439H1 SATMON017 g2243115 BLASTN 448 1e−43 76 871-700266615 700266615H1 SATMON017 g2243116 BLASTX 179 1e−17 73 872-700342655 700342655H1 SATMON021 g2257743 BLASTX 213 1e−22 82 873-700343285 700343285H1 SATMON021 g2257742 BLASTN 437 1e−25 66 874-700467533 700467533H1 SATMON025 g2243115 BLASTN 439 1e−26 79 875-700548678 700548678H1 SATMON022 g147979 BLASTX 147 1e−13 62 876-700613618 700613618H1 SATMON033 g500850 BLASTN 965 1e−81 98 877-L30691987 LIB3069-018-Q1-K1-A3 LIB3069 g2243115 BLASTN 266 1e−10 57 87812201 700457103H1 SATMON029 g2257742 BLASTN 789 1e−56 76 879 12201700457111H1 SATMON029 g2243115 BLASTN 513 1e−33 76 880 12931 700380864H1SATMON023 g500852 BLASTN 1450 1e−111 95 881 12931 700105610H1 SATMON010g500852 BLASTN 1046 1e−105 95 882 12931 700380848H1 SATMON023 g500852BLASTN 1193 1e−96 95 883 12931 700205392H1 SATMON003 g500852 BLASTN 10651e−94 93 884 12931 700552314H1 SATMON022 g500852 BLASTN 908 1e−86 90 88512931 700551915H1 SATMON022 g500852 BLASTN 1090 1e−81 90 886 16037700344509H1 SATMON021 g500852 BLASTN 1184 1e−89 92 887 16037 700345170H1SATMON021 g500852 BLASTN 600 1e−58 85 888 16157 700212607H1 SATMON016g2243115 BLASTN 914 1e−67 76 889 16157 700094809H1 SATMON008 g2243116BLASTX 144 1e−12 84 890 19231 700091761H1 SATMON011 g500850 BLASTN 13581e−104 98 891 19231 700612568H1 SATMON033 g500850 BLASTN 1137 1e−102 99892 22303 700553291H1 SATMON022 g2243115 BLASTN 644 1e−44 70 893 22303700553382H1 SATMON022 g2243115 BLASTN 418 1e−39 70 894 28000LIB143-061-Q1-E1-C7 LIB143 g2243115 BLASTN 1132 1e−85 75 895 28000700474110H1 SATMON025 g2243115 BLASTN 509 1e−33 75 896 30401 700620948H1SATMON034 g500852 BLASTN 327 1e−30 88 897 32907 LIB143-038-Q1-E1-B11LIB143 g500850 BLASTN 1864 1e−146 96 898 32907 700096779H1 SATMON008g500850 BLASTN 1490 1e−115 97 899 5616 700346488H1 SATMON021 g2243115BLASTN 664 1e−46 71 900 5616 700196138H1 SATMON014 g2243115 BLASTN 6301e−43 74 2624 -700556108 700556108H1 SOYMON001 g2243115 BLASTN 700 1e−4974 2625 -700663367 700663367H1 SOYMON005 g2243115 BLASTN 737 1e−52 772626 -700733301 700733301H1 SOYMON010 g2243115 BLASTN 751 1e−53 78 2627-700747979 700747979H1 SOYMON013 g2257742 BLASTN 449 1e−27 70 2628-700832664 700832664H1 SOYMON019 g167547 BLASTN 322 1e−44 78 2629-700843925 700843925H1 SOYMON021 g167547 BLASTN 616 1e−42 71 2630-700888516 700888516H1 SOYMON024 g464225 BLASTX 193 1e−19 78 2631-700892002 700892002H1 SOYMON024 g2243115 BLASTN 363 1e−21 79 2632-700959057 700959057H1 SOYMON022 g2257742 BLASTN 497 1e−32 72 2633-700971891 700971891H1 SOYMON005 g167547 BLASTN 699 1e−49 74 2634-700984812 700984812H1 SOYMON009 g2257742 BLASTN 801 1e−57 77 2635-701069254 701069254H1 SOYMON034 g2243115 BLASTN 260 1e−23 75 2636-701120341 701120341H1 SOYMON037 g2243115 BLASTN 567 1e−38 77 2637-GM35173 LIB3051-037-Q1-K1-B9 LIB3051 g2970554 BLASTN 193 1e−11 83 263815020 700557507H1 SOYMON001 g167547 BLASTN 914 1e−67 79 2639 15020700666142H1 SOYMON005 g1107460 BLASTN 767 1e−55 77 2640 18237700797368H1 SOYMON017 g2257742 BLASTN 819 1e−59 81 2641 18237700797360H1 SOYMON017 g2257742 BLASTN 809 1e−58 84 2642 19332LIB3056-004-Q1-N1-D5 LIB3056 g2243115 BLASTN 1118 1e−84 74 2643 19332700786255H2 SOYMON011 g2257742 BLASTN 626 1e−43 72 2644 19332700684751H1 SOYMON008 g2257742 BLASTN 583 1e−39 73 2645 21954701100440H1 SOYMON028 g167547 BLASTN 835 1e−60 78 2646 21954 701059173H1SOYMON033 g167547 BLASTN 719 1e−51 75 2647 26336 701003103H1 SOYMON019g2243115 BLASTN 877 1e−64 79 2648 26336 700976874H1 SOYMON009 g2243115BLASTN 880 1e−64 79

ASPARTATE-SEMIALDEHYDE DEHYDROGENASE (EC 1.2.1.11) Seq No. Cluster IDCloneID Library NCBI gi Method Score P-value % Ident 901 -700439614700439614H1 SATMON026 g2314350 BLASTX 107 1e−13 52 902 -701183695701183695H1 SATMONN06 g289910 BLASTX 150 1e−13 69 903 -L1487398LIB148-064-Q1-E1-D10 LIB148 g1749466 BLASTX 178 1e−33 55 904 -L30622830LIB3062-028-Q1-K1-A8 LIB3062 g1085109 BLASTX 108 1e−40 48 2649-700756763 700756763H1 SOYMON014 g1359593 BLASTX 71 1e−8 52 2650-700830054 700830054H1 SOYMON019 g142828 BLASTX 87 1e−10 41 2651-701105617 701105617H1 SOYMON036 g142828 BLASTX 215 1e−22 57 2652-GM8539 LIB3039-047-Q1-E1-C6 LIB3039 g142828 BLASTX 188 1e−35 50 265330187 LIB3049-001-Q1-E1-F2 LIB3049 g1359593 BLASTX 136 1e−26 54 265430187 700556105H1 SOYMON001 g1359593 BLASTX 132 1e−11 56

O-SUCCINYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.9) Seq No. Cluster IDCloneID Library NCBI gi Method Score P-value % Ident 905 -700049526700049526H1 SATMON003 g2198852 BLASTN 251 1e−9 76 906 -700086788700086788H1 SATMON011 g2198850 BLASTN 631 1e−61 88 907 -700460589700460589H1 SATMON030 g2198850 BLASTN 251 1e−32 83 908 -700577561700577561H1 SATMON031 g2198852 BLASTN 207 1e−15 82 909 -700579840700579840H1 SATMON031 g2198850 BLASTN 216 1e−12 94 910 -700616013700616013H1 SATMON033 g2198852 BLASTN 278 1e−39 81 911 -L1892785LIB189-011-Q1-E1-A10 LIB189 g2198852 BLASTN 311 1e−14 82 912 -L30594402LIB3059-042-Q1-K1-E11 LIB3059 g2198852 BLASTN 489 1e−40 84 913-L30604293 LIB3060-028-Q1-K1-E7 LIB3060 g2198852 BLASTN 380 1e−20 83 914-L30693289 LIB3069-016-Q1-K1-G10 LIB3069 g2198852 BLASTN 197 1e−10 79915 10571 700224881H1 SATMON011 g2198852 BLASTN 319 1e−15 75 916 10801700429049H1 SATMONN01 g2198852 BLASTN 210 1e−16 82 917 10801 700167813H1SATMON013 g2198852 BLASTN 200 1e−15 82 918 10801 700074027H1 SATMON007g2198852 BLASTN 178 1e−11 80 919 16379 LIB3061-035-Q1-K1-B6 LIB3061g2198850 BLASTN 2184 1e−173 99 920 16379 700259309H1 SATMON017 g2198850BLASTN 1259 1e−108 92 921 16379 700051696H1 SATMON003 g2198850 BLASTN1392 1e−107 96 922 16379 700239553H1 SATMON010 g2198850 BLASTN 12651e−96 96 923 16379 700042846H1 SATMON004 g2198850 BLASTN 1208 1e−91 95924 16379 700092741H1 SATMON008 g2198850 BLASTN 1042 1e−89 95 925 16379LIB3061-017-Q1-K1-D7 LIB3061 g2198850 BLASTN 1183 1e−89 99 926 16379700206765H1 SATMON003 g2198850 BLASTN 1095 1e−82 81 927 16379700150281H1 SATMON007 g2198850 BLASTN 1024 1e−76 94 928 16379700165862H1 SATMON013 g2198850 BLASTN 948 1e−75 94 929 2221LIB3060-017-Q1-K1-B10 LIB3060 g2198850 BLASTN 1819 1e−161 99 930 2221LIB84-006-Q1-E1-F3 LIB84 g2198852 BLASTN 1615 1e−153 97 931 2221700575334H1 SATMON030 g2198850 BLASTN 1570 1e−124 99 932 2221700206250H1 SATMON003 g2198850 BLASTN 1515 1e−117 100 933 2221700095073H1 SATMON008 g2198852 BLASTN 1490 1e−115 100 934 2221700571230H1 SATMON030 g2198850 BLASTN 1355 1e−114 93 935 2221700157358H1 SATMON012 g2198850 BLASTN 1370 1e−105 100 936 2221700379811H1 SATMON021 g2198850 BLASTN 1345 1e−103 93 937 2221700041570H1 SATMON004 g2198850 BLASTN 1325 1e−101 100 938 2221700104063H1 SATMON010 g2198850 BLASTN 1157 1e−95 90 939 2221 700378265H1SATMON019 g2198850 BLASTN 771 1e−94 99 940 2221 700235329H1 SATMON010g2198850 BLASTN 1234 1e−94 93 941 2221 LIB3068-057-Q1-K1-D1 LIB3068g2198852 BLASTN 1209 1e−91 94 942 2221 700159158H1 SATMON012 g2198850BLASTN 1174 1e−89 94 943 2221 700580854H1 SATMON031 g2198850 BLASTN 7541e−86 92 944 2221 700623409H1 SATMON034 g2198850 BLASTN 910 1e−84 96 9452221 701164706H1 SATMONN04 g2198852 BLASTN 577 1e−83 94 946 2221700164719H1 SATMON013 g2198852 BLASTN 831 1e−83 99 947 2221 700158146H1SATMON012 g2198850 BLASTN 1100 1e−82 93 948 2221 700203970H1 SATMON003g2198852 BLASTN 996 1e−80 99 949 2221 700158313H1 SATMON012 g2198850BLASTN 1036 1e−77 93 950 2221 700425211H1 SATMONN01 g2198852 BLASTN 4851e−61 96 951 2221 700167764H1 SATMON013 g2198850 BLASTN 793 1e−57 93 95223788 700102780H1 SATMON010 g2198852 BLASTN 1131 1e−104 99 953 23788701167458H1 SATMONN05 g2198852 BLASTN 1069 1e−90 93 2655 -700900206700900206H1 SOYMON027 g1742961 BLASTX 215 1e−24 78 2656 -GM40351LIB3051-114-Q1-K1-H12 LIB3051 g2198851 BLASTX 122 1e−25 96 2657 12502701101592H1 SOYMON028 g146846 BLASTX 103 1e−18 44 2658 12502 701106834H1SOYMON036 g146846 BLASTX 103 1e−18 44 2659 13820 LIB3055-003-Q1-N1-F12LIB3055 g3202028 BLASTX 193 1e−35 94 2660 8119 700989656H1 SOYMON011g1742960 BLASTN 815 1e−59 79

CYSTATHIONINE β-LYASE (EC 4.4.1.8) Seq No. Cluster ID CloneID LibraryNCBI gi Method Score P-value % Ident 954 -700155172 700155172H1SATMON007 g704397 BLASTX 361 1e−43 78 955 -L362943 LIB36-013-Q1-E1-G10LIB36 g704396 BLASTN 818 1e−59 75 956 19856 700240664H1 SATMON010g704396 BLASTN 496 1e−31 69 957 19856 700572496H1 SATMON030 g704396BLASTN 446 1e−26 68 958 22960 701170964H1 SATMONN05 g704396 BLASTN 7781e−56 78 959 22960 LIB3061-002-Q1-K2-F9 LIB3061 g704396 BLASTN 765 1e−5377 960 22960 701172780H2 SATMONN05 g704396 BLASTN 686 1e−48 73 961 22960700578571H1 SATMON031 g704397 BLASTX 222 1e−23 89 962 30752LIB3078-055-Q1-K1-C8 LIB3078 g704396 BLASTN 747 1e−51 71 963 30752700086603H1 SATMON011 g704397 BLASTX 192 1e−18 64 2661 -701001147701001147H1 SOYMON018 g704396 BLASTN 847 1e−61 78 2662 18602 700566066H1SOYMON002 g704396 BLASTN 751 1e−57 79 2663 18602 700890955H1 SOYMON024g704396 BLASTN 698 1e−49 77 2664 18602 700896865H1 SOYMON027 g704396BLASTN 682 1e−48 77 2665 5144 LIB3050-006-Q1-E1-A9 LIB3050 g1399263BLASTX 96 1e−31 41

5-METHYLTETRAHYDROPTEROYLTRIGLUTAMATE - HOMOCYSTEINE S-METHYLTRANSFERASE(EC 2.1.1.14) Seq No. Cluster ID CloneID Library NCBI gi Method ScoreP-value % Ident 964 -700212217 700212217H1 SATMON016 g886471 BLASTX 1511e−16 93 965 -700333966 700333966H1 SATMON019 g886470 BLASTN 370 1e−2366 966 -700377403 700377403H1 SATMON019 g2738247 BLASTN 305 1e−26 80 967-700571893 700571893H1 SATMON030 g974781 BLASTN 262 1e−20 80 968-701165656 701165656H1 SATMONN04 g2738247 BLASTN 450 1e−50 73 969-L30622954 LIB3062-030-Q1-K1-C9 LIB3062 g886470 BLASTN 366 1e−26 81 970-L30662004 LIB3066-026-Q1-K1-F1 LIB3066 g2738248 BLASTX 140 1e−28 77 971-L30671450 LIB3067-001-Q1-K1-C6 LIB3067 g2738247 BLASTN 553 1e−46 69 972-L30694410 LIB3069-057-Q1-K1-B4 LIB3069 g886470 BLASTN 447 1e−26 67 9731 700452404H1 SATMON028 g453939 BLASTX 59 1e−17 94 974 13513 700049012H1SATMON003 g1814402 BLASTN 1006 1e−74 81 975 13513 700086058H1 SATMON011g1814402 BLASTN 982 1e−72 81 976 13513 700235814H1 SATMON010 g1814402BLASTN 890 1e−65 79 977 13513 700170015H1 SATMON013 g1814402 BLASTN 5781e−39 78 978 3835 700093161H1 SATMON008 g974781 BLASTN 858 1e−62 76 9793835 700223321H1 SATMON011 g974781 BLASTN 836 1e−60 80 980 3835700454003H1 SATMON029 g974781 BLASTN 823 1e−59 82 981 3835 700238925H1SATMON010 g2738247 BLASTN 733 1e−55 74 982 3835 700075142H1 SATMON007g2738247 BLASTN 569 1e−52 73 983 3835 700151969H1 SATMON007 g974781BLASTN 660 1e−46 76 984 3835 700281434H2 SATMON019 g974781 BLASTN 5601e−45 79 985 3835 700084914H1 SATMON011 g974781 BLASTN 440 1e−36 79 9863835 700215135H1 SATMON016 g974781 BLASTN 543 1e−36 78 987 3835700202218H1 SATMON003 g974781 BLASTN 389 1e−23 78 988 3835 700281467H2SATMON019 g2738248 BLASTX 123 1e−12 71 989 456 LIB3059-019-Q1-K1-G6LIB3059 g974781 BLASTN 1493 1e−115 83 990 456 LIB3068-012-Q1-K1-G7LIB3068 g974781 BLASTN 1471 1e−113 81 991 456 LIB3061-050-Q1-K1-G10LIB3061 g886470 BLASTN 1285 1e−98 82 992 456 LIB3067-043-Q1-K1-F9LIB3067 g2738247 BLASTN 1287 1e−98 79 993 456 700087169H1 SATMON011g1814402 BLASTN 1245 1e−94 86 994 456 LIB3069-030-Q1-K1-G9 LIB3069g1814402 BLASTN 772 1e−93 80 995 456 LIB3069-019-Q1-K1-G11 LIB3069g1814402 BLASTN 1202 1e−91 81 996 456 700202514H1 SATMON003 g1814402BLASTN 1134 1e−89 84 997 456 LIB3062-011-Q1-K1-F2 LIB3062 g886470 BLASTN664 1e−86 84 998 456 LIB143-015-Q1-E1-A1 LIB143 g974781 BLASTN 11431e−86 81 999 456 700570326H1 SATMON030 g974781 BLASTN 1089 1e−85 83 1000456 700103727H1 SATMON010 g2738247 BLASTN 1132 1e−85 86 1001 456700574823H1 SATMON030 g1814402 BLASTN 1135 1e−85 80 1002 456 700091666H1SATMON011 g1814402 BLASTN 1136 1e−85 84 1003 456 LIB3069-033-Q1-K1-E9LIB3069 g2738247 BLASTN 1115 1e−84 79 1004 456 700572634H1 SATMON030g1814402 BLASTN 1123 1e−84 85 1005 456 700211829H1 SATMON016 g2738247BLASTN 1096 1e−82 84 1006 456 700087109H1 SATMON011 g1814402 BLASTN 11021e−82 85 1007 456 LIB3059-003-Q1-K1-B1 LIB3059 g1814402 BLASTN 961 1e−8182 1008 456 700047556H1 SATMON003 g2738247 BLASTN 976 1e−81 82 1009 456700201925H1 SATMON003 g2738247 BLASTN 1001 1e−81 84 1010 456 700209343H1SATMON016 g1814402 BLASTN 1081 1e−81 82 1011 456 700348416H1 SATMON023g1814402 BLASTN 1081 1e−81 83 1012 456 700095559H1 SATMON008 g1814402BLASTN 1085 1e−81 82 1013 456 700073472H1 SATMON007 g1814402 BLASTN 10891e−81 82 1014 456 LIB3062-057-Q1-K1-C2 LIB3062 g886470 BLASTN 1073 1e−8078 1015 456 700093445H1 SATMON008 g1814402 BLASTN 1076 1e−80 84 1016 456LIB143-026-Q1-E1-H3 LIB143 g2738247 BLASTN 648 1e−79 79 1017 456700206240H1 SATMON003 g1814402 BLASTN 1057 1e−79 82 1018 456 700091907H1SATMON011 g886470 BLASTN 1062 1e−79 80 1019 456 700258178H1 SATMON017g1814402 BLASTN 1064 1e−79 84 1020 456 700086565H1 SATMON011 g1814402BLASTN 1065 1e−79 82 1021 456 700471452H1 SATMON025 g2738247 BLASTN 10451e−78 85 1022 456 700243858H1 SATMON010 g1814402 BLASTN 1048 1e−78 851023 456 700352224H1 SATMON023 g2738247 BLASTN 1051 1e−78 84 1024 456700084769H1 SATMON011 g1814402 BLASTN 1054 1e−78 83 1025 456 700082818H1SATMON011 g886470 BLASTN 984 1e−77 84 1026 456 700331974H1 SATMON019g886470 BLASTN 1031 1e−77 82 1027 456 700086203H1 SATMON011 g1814402BLASTN 1032 1e−77 81 1028 456 700075826H1 SATMON007 g2738247 BLASTN 10381e−77 82 1029 456 700104555H1 SATMON010 g886470 BLASTN 1039 1e−77 791030 456 700074084H1 SATMON007 g2738247 BLASTN 1039 1e−77 87 1031 456700076872H1 SATMON007 g1814402 BLASTN 700 1e−76 83 1032 456 700050704H1SATMON003 g1814402 BLASTN 880 1e−76 82 1033 456 700030215H1 SATMON003g974781 BLASTN 1019 1e−76 83 1034 456 700077416H1 SATMON007 g886470BLASTN 1019 1e−76 79 1035 456 700093982H1 SATMON008 g1814402 BLASTN 10191e−76 82 1036 456 700048847H1 SATMON003 g2738247 BLASTN 1023 1e−76 851037 456 700090739H1 SATMON011 g886470 BLASTN 1026 1e−76 82 1038 456700092904H1 SATMON008 g886470 BLASTN 1026 1e−76 82 1039 456LIB3068-009-Q1-K1-E5 LIB3068 g1814402 BLASTN 1027 1e−76 73 1040 456700336705H1 SATMON019 g1814402 BLASTN 747 1e−75 83 1041 456 700344872H1SATMON021 g1814402 BLASTN 768 1e−75 85 1042 456 LIB3066-019-Q1-K1-E1LIB3066 g886470 BLASTN 868 1e−75 81 1043 456 700086322H1 SATMON011g2738247 BLASTN 1015 1e−75 81 1044 456 LIB3067-052-Q1-K1-B12 LIB3067g974781 BLASTN 1033 1e−75 81 1045 456 700221395H1 SATMON011 g2738247BLASTN 1000 1e−74 85 1046 456 700092483H1 SATMON008 g1814402 BLASTN 10011e−74 82 1047 456 700618708H1 SATMON034 g886470 BLASTN 1003 1e−74 821048 456 700074191H1 SATMON007 g2738247 BLASTN 1005 1e−74 86 1049 456LIB143-055-Q1-E1-F10 LIB143 g1814402 BLASTN 986 1e−73 83 1050 456700025955H1 SATMON003 g1814402 BLASTN 993 1e−73 84 1051 456 700574824H1SATMON030 g886470 BLASTN 564 1e−72 80 1052 456 700201527H1 SATMON003g974781 BLASTN 812 1e−72 81 1053 456 700092202H1 SATMON008 g886470BLASTN 946 1e−72 79 1054 456 700223674H1 SATMON011 g2738247 BLASTN 9711e−72 83 1055 456 700092638H1 SATMON008 g1814402 BLASTN 972 1e−72 841056 456 700090025H1 SATMON011 g974781 BLASTN 973 1e−72 81 1057 456700217069H1 SATMON016 g2738247 BLASTN 976 1e−72 83 1058 456 700349142H1SATMON023 g1814402 BLASTN 980 1e−72 82 1059 456 700088387H1 SATMON011g2738247 BLASTN 980 1e−72 85 1060 456 700215102H1 SATMON016 g886470BLASTN 980 1e−72 79 1061 456 700456708H1 SATMON029 g2738247 BLASTN 9821e−72 82 1062 456 700210025H1 SATMON016 g1814402 BLASTN 982 1e−72 811063 456 700335345H1 SATMON019 g974781 BLASTN 982 1e−72 84 1064 456700085257H1 SATMON011 g1814402 BLASTN 554 1e−71 82 1065 456 700026781H1SATMON003 g974781 BLASTN 961 1e−71 82 1066 456 700224265H1 SATMON011g2738247 BLASTN 962 1e−71 83 1067 456 700083282H1 SATMON011 g1814402BLASTN 962 1e−71 83 1068 456 700381324H1 SATMON023 g1814402 BLASTN 9651e−71 82 1069 456 700212017H1 SATMON016 g1814402 BLASTN 640 1e−70 831070 456 700088053H1 SATMON011 g886470 BLASTN 853 1e−70 84 1071 456LIB3062-032-Q1-K1-C2 LIB3062 g1814402 BLASTN 883 1e−70 80 1072 456700083030H1 SATMON011 g1814402 BLASTN 948 1e−70 81 1073 456 700613980H1SATMON033 g1814402 BLASTN 950 1e−70 83 1074 456 700347019H1 SATMON021g974781 BLASTN 952 1e−70 81 1075 456 700335621H1 SATMON019 g1814402BLASTN 952 1e−70 80 1076 456 700094101H1 SATMON008 g1814402 BLASTN 9531e−70 83 1077 456 700073128H1 SATMON007 g2738247 BLASTN 954 1e−70 781078 456 700076245H1 SATMON007 g1814402 BLASTN 955 1e−70 83 1079 456700071903H1 SATMON007 g886470 BLASTN 956 1e−70 81 1080 456 700090035H1SATMON011 g886470 BLASTN 935 1e−69 80 1081 456 700405323H1 SATMON029g1814402 BLASTN 937 1e−69 83 1082 456 700050121H1 SATMON003 g886470BLASTN 938 1e−69 83 1083 456 700224714H1 SATMON011 g1814402 BLASTN 9391e−69 82 1084 456 700281535H2 SATMON019 g1814402 BLASTN 940 1e−69 811085 456 700222093H1 SATMON011 g2738247 BLASTN 941 1e−69 81 1086 456700094659H1 SATMON008 g1814402 BLASTN 942 1e−69 84 1087 456 700237187H1SATMON010 g1814402 BLASTN 942 1e−69 84 1088 456 700023210H1 SATMON003g1814402 BLASTN 943 1e−69 83 1089 456 700072813H1 SATMON007 g1814402BLASTN 944 1e−69 82 1090 456 700075720H1 SATMON007 g1814402 BLASTN 9461e−69 83 1091 456 700072107H1 SATMON007 g1814402 BLASTN 946 1e−69 801092 456 700082225H1 SATMON011 g1814402 BLASTN 691 1e−68 82 1093 456700085411H1 SATMON011 g1814402 BLASTN 723 1e−68 83 1094 456 700454357H1SATMON029 g2738247 BLASTN 926 1e−68 82 1095 456 700620976H1 SATMON034g1814402 BLASTN 927 1e−68 81 1096 456 700241483H1 SATMON010 g1814402BLASTN 931 1e−68 86 1097 456 700355459H1 SATMON024 g974781 BLASTN 9311e−68 82 1098 456 700095875H1 SATMON008 g1814402 BLASTN 931 1e−68 871099 456 700076702H1 SATMON007 g1814402 BLASTN 934 1e−68 84 1100 456700212538H1 SATMON016 g1814402 BLASTN 689 1e−67 85 1101 456 700086479H1SATMON011 g1814402 BLASTN 715 1e−67 82 1102 456 700453684H1 SATMON028g1814402 BLASTN 747 1e−67 83 1103 456 700204454H1 SATMON003 g1814402BLASTN 797 1e−67 82 1104 456 700612577H1 SATMON033 g974781 BLASTN 9111e−67 81 1105 456 700474018H1 SATMON025 g886470 BLASTN 917 1e−67 81 1106456 700213602H1 SATMON016 g886470 BLASTN 917 1e−67 76 1107 456700076911H1 SATMON007 g974781 BLASTN 917 1e−67 81 1108 456 700223313H1SATMON011 g886470 BLASTN 919 1e−67 84 1109 456 700220692H1 SATMON011g974781 BLASTN 920 1e−67 81 1110 456 700258038H1 SATMON017 g886470BLASTN 920 1e−67 83 1111 456 700095862H1 SATMON008 g1814402 BLASTN 9221e−67 86 1112 456 700213396H1 SATMON016 g886470 BLASTN 922 1e−67 83 1113456 700354259H1 SATMON024 g974781 BLASTN 638 1e−66 81 1114 456700471505H1 SATMON025 g886470 BLASTN 681 1e−66 83 1115 456 700202439H1SATMON003 g1814402 BLASTN 706 1e−66 82 1116 456 700223609H1 SATMON011g974781 BLASTN 770 1e−66 83 1117 456 700405260H1 SATMON028 g1814402BLASTN 782 1e−66 86 1118 456 700085214H1 SATMON011 g886470 BLASTN 8141e−66 84 1119 456 700211194H1 SATMON016 g1814402 BLASTN 902 1e−66 841120 456 700106641H1 SATMON010 g1814402 BLASTN 903 1e−66 82 1121 456700405170H1 SATMON028 g974781 BLASTN 904 1e−66 79 1122 456 700090015H1SATMON011 g886470 BLASTN 905 1e−66 80 1123 456 700623154H1 SATMON034g886470 BLASTN 906 1e−66 81 1124 456 700458895H1 SATMON029 g886470BLASTN 906 1e−66 83 1125 456 700094073H1 SATMON008 g886470 BLASTN 9071e−66 78 1126 456 700158508H1 SATMON012 g2738247 BLASTN 909 1e−66 821127 456 700027366H1 SATMON003 g1814402 BLASTN 910 1e−66 81 1128 456700073636H1 SATMON007 g1814402 BLASTN 615 1e−65 87 1129 456 700220207H1SATMON011 g886470 BLASTN 785 1e−65 84 1130 456 LIB143-007-Q1-E1-A6LIB143 g886470 BLASTN 807 1e−65 80 1131 456 700575288H1 SATMON030g1814402 BLASTN 887 1e−65 83 1132 456 700219870H1 SATMON011 g974781BLASTN 887 1e−65 82 1133 456 701184573H1 SATMONN06 g974781 BLASTN 8881e−65 81 1134 456 700076946H1 SATMON007 g1814402 BLASTN 891 1e−65 861135 456 700214808H1 SATMON016 g2738247 BLASTN 893 1e−65 81 1136 456700096274H1 SATMON008 g1814402 BLASTN 894 1e−65 80 1137 456 700160445H1SATMON012 g1814402 BLASTN 896 1e−65 85 1138 456 700071727H1 SATMON007g974781 BLASTN 898 1e−65 80 1139 456 700571751H1 SATMON030 g1814402BLASTN 898 1e−65 78 1140 456 700215107H1 SATMON016 g2738247 BLASTN 5071e−64 77 1141 456 LIB3067-057-Q1-K1-C6 LIB3067 g2738247 BLASTN 555 1e−6476 1142 456 700614743H1 SATMON033 g886470 BLASTN 748 1e−64 81 1143 456700444971H1 SATMON027 g974781 BLASTN 803 1e−64 83 1144 456 700224573H1SATMON011 g1814402 BLASTN 805 1e−64 86 1145 456 700019276H1 SATMON001g1814402 BLASTN 875 1e−64 83 1146 456 700210440H1 SATMON016 g2738247BLASTN 881 1e−64 80 1147 456 700093168H1 SATMON008 g1814402 BLASTN 8851e−64 86 1148 456 700224478H1 SATMON011 g886470 BLASTN 886 1e−64 81 1149456 700261958H1 SATMON017 g1814402 BLASTN 781 1e−63 79 1150 456700074221H1 SATMON007 g1814402 BLASTN 825 1e−63 81 1151 456 700089966H1SATMON011 g1814402 BLASTN 863 1e−63 84 1152 456 700210422H1 SATMON016g1814402 BLASTN 863 1e−63 81 1153 456 700160272H1 SATMON012 g1814402BLASTN 863 1e−63 85 1154 456 700444282H1 SATMON027 g2738247 BLASTN 5491e−62 84 1155 456 700206489H1 SATMON003 g2738247 BLASTN 597 1e−62 801156 456 700446452H1 SATMON027 g974781 BLASTN 711 1e−62 80 1157 456700265473H1 SATMON017 g1814402 BLASTN 715 1e−62 79 1158 456 700473708H1SATMON025 g1814402 BLASTN 762 1e−62 82 1159 456 LIB3069-025-Q1-K1-A7LIB3069 g886470 BLASTN 852 1e−62 80 1160 456 700166631H1 SATMON013g2738247 BLASTN 858 1e−62 85 1161 456 700582413H1 SATMON031 g1814402BLASTN 861 1e−62 78 1162 456 700071904H1 SATMON007 g2738247 BLASTN 5441e−61 77 1163 456 700622655H1 SATMON034 g1814402 BLASTN 592 1e−61 821164 456 700082541H1 SATMON011 g1814402 BLASTN 793 1e−61 82 1165 456700335378H1 SATMON019 g2738247 BLASTN 841 1e−61 76 1166 456 700203585H1SATMON003 g2738247 BLASTN 846 1e−61 86 1167 456 700053043H1 SATMON007g2738247 BLASTN 847 1e−61 81 1168 456 700163654H1 SATMON013 g1814402BLASTN 474 1e−60 84 1169 456 700352269H1 SATMON023 g1814402 BLASTN 4791e−60 84 1170 456 700106187H1 SATMON010 g1814402 BLASTN 631 1e−60 811171 456 LIB143-007-Q1-E1-A7 LIB143 g886470 BLASTN 759 1e−60 76 1172 456700458694H1 SATMON029 g1814402 BLASTN 827 1e−60 86 1173 456 700213509H1SATMON016 g886470 BLASTN 834 1e−60 81 1174 456 700150191H1 SATMON007g2738247 BLASTN 834 1e−60 82 1175 456 700157594H1 SATMON012 g1814402BLASTN 835 1e−60 80 1176 456 700094046H1 SATMON008 g974781 BLASTN 8361e−60 79 1177 456 700220254H1 SATMON011 g2738247 BLASTN 687 1e−59 771178 456 700095485H1 SATMON008 g886470 BLASTN 819 1e−59 77 1179 456LIB143-006-Q1-E1-B3 LIB143 g1814402 BLASTN 830 1e−59 65 1180 456700071763H1 SATMON007 g2738247 BLASTN 565 1e−58 75 1181 456 700082259H1SATMON011 g1814402 BLASTN 707 1e−58 84 1182 456 700020717H1 SATMON001g886470 BLASTN 805 1e−58 81 1183 456 700041892H1 SATMON004 g1814402BLASTN 805 1e−58 86 1184 456 700356573H1 SATMON024 g1814402 BLASTN 8111e−58 81 1185 456 700171632H1 SATMON013 g2738247 BLASTN 811 1e−58 851186 456 700222516H1 SATMON011 g1814402 BLASTN 812 1e−58 87 1187 456700466679H1 SATMON025 g1814402 BLASTN 812 1e−58 87 1188 456 700208929H1SATMON016 g1814402 BLASTN 407 1e−57 78 1189 456 700355014H1 SATMON024g1814402 BLASTN 592 1e−57 85 1190 456 701159834H1 SATMONN04 g1814402BLASTN 683 1e−57 77 1191 456 700351137H1 SATMON023 g974781 BLASTN 7251e−57 79 1192 456 700027308H1 SATMON003 g2738247 BLASTN 791 1e−57 801193 456 700142587H1 SATMON012 g974781 BLASTN 791 1e−57 83 1194 456700170157H1 SATMON013 g1814402 BLASTN 791 1e−57 81 1195 456 700019438H1SATMON001 g974781 BLASTN 798 1e−57 82 1196 456 700166430H1 SATMON013g886470 BLASTN 799 1e−57 84 1197 456 700047369H1 SATMON003 g886470BLASTN 799 1e−57 80 1198 456 700152373H1 SATMON007 g2738247 BLASTN 7991e−57 84 1199 456 700050824H1 SATMON003 g2738247 BLASTN 605 1e−56 811200 456 700204588H1 SATMON003 g886470 BLASTN 720 1e−56 82 1201 456700071667H1 SATMON007 g1814402 BLASTN 787 1e−56 85 1202 456 701185269H1SATMONN06 g886470 BLASTN 598 1e−55 76 1203 456 700243844H1 SATMON010g2738247 BLASTN 769 1e−55 79 1204 456 700221969H1 SATMON011 g974781BLASTN 771 1e−55 79 1205 456 700335706H1 SATMON019 g1814402 BLASTN 7731e−55 80 1206 456 700622285H1 SATMON034 g1814402 BLASTN 774 1e−55 821207 456 700089665H1 SATMON011 g2738247 BLASTN 774 1e−55 78 1208 456700464833H1 SATMON025 g2738247 BLASTN 455 1e−54 72 1209 456LIB3069-057-Q1-K1-A6 LIB3069 g974781 BLASTN 564 1e−54 76 1210 456700156818H1 SATMON012 g974781 BLASTN 757 1e−54 81 1211 456 700219420H1SATMON011 g974781 BLASTN 758 1e−54 79 1212 456 700152570H1 SATMON007g1814402 BLASTN 761 1e−54 86 1213 456 700084484H1 SATMON011 g1814402BLASTN 764 1e−54 84 1214 456 700152208H1 SATMON007 g1814402 BLASTN 7651e−54 82 1215 456 700550889H1 SATMON022 g2738247 BLASTN 546 1e−53 761216 456 700025869H1 SATMON003 g2738247 BLASTN 675 1e−53 79 1217 456700383075H1 SATMON024 g974781 BLASTN 724 1e−53 81 1218 456 700575104H1SATMON030 g1814402 BLASTN 745 1e−53 70 1219 456 LIB3069-022-Q1-K1-C1LIB3069 g974781 BLASTN 768 1e−53 81 1220 456 700215272H1 SATMON016g1814402 BLASTN 674 1e−52 83 1221 456 700167881H1 SATMON013 g974781BLASTN 737 1e−52 79 1222 456 700019727H1 SATMON001 g2738247 BLASTN 7371e−52 81 1223 456 700351776H1 SATMON023 g886470 BLASTN 741 1e−52 76 1224456 700210543H1 SATMON016 g974781 BLASTN 742 1e−52 85 1225 456700571604H1 SATMON030 g1814402 BLASTN 360 1e−51 76 1226 456 700169777H1SATMON013 g1814402 BLASTN 426 1e−51 83 1227 456 700096744H1 SATMON008g2738247 BLASTN 539 1e−51 79 1228 456 700383157H1 SATMON024 g974781BLASTN 667 1e−51 79 1229 456 700155764H1 SATMON007 g886470 BLASTN 7211e−51 75 1230 456 700150817H1 SATMON007 g1814402 BLASTN 727 1e−51 751231 456 700619660H1 SATMON034 g886470 BLASTN 727 1e−51 73 1232 456700471085H1 SATMON025 g1814402 BLASTN 406 1e−50 79 1233 456 700163226H1SATMON013 g2738247 BLASTN 475 1e−50 78 1234 456 700444567H1 SATMON027g1814402 BLASTN 498 1e−50 81 1235 456 700457303H1 SATMON029 g2738247BLASTN 662 1e−50 80 1236 456 700088065H1 SATMON011 g886470 BLASTN 7071e−50 79 1237 456 700152592H1 SATMON007 g886470 BLASTN 716 1e−50 80 1238456 LIB3068-061-Q1-K1-B2 LIB3068 g886470 BLASTN 730 1e−50 78 1239 456700331880H1 SATMON019 g1814402 BLASTN 697 1e−49 85 1240 456 700218025H1SATMON016 g886470 BLASTN 700 1e−49 79 1241 456 700348643H1 SATMON023g1814402 BLASTN 701 1e−49 80 1242 456 700236975H1 SATMON010 g974781BLASTN 704 1e−49 81 1243 456 700442968H1 SATMON026 g886470 BLASTN 4181e−48 79 1244 456 700479529H1 SATMON034 g2738247 BLASTN 508 1e−48 751245 456 700160983H1 SATMON012 g1814402 BLASTN 690 1e−48 83 1246 456700050185H1 SATMON003 g974781 BLASTN 421 1e−47 75 1247 456 700611764H1SATMON022 g1814402 BLASTN 503 1e−47 79 1248 456 700622669H1 SATMON034g1814402 BLASTN 549 1e−47 79 1249 456 700153237H1 SATMON007 g1814402BLASTN 672 1e−47 80 1250 456 700242703H1 SATMON010 g886470 BLASTN 6821e−47 81 1251 456 700165177H1 SATMON013 g2738247 BLASTN 682 1e−47 781252 456 700379889H1 SATMON021 g886470 BLASTN 682 1e−47 85 1253 456700449243H1 SATMON028 g1814402 BLASTN 357 1e−46 83 1254 456 700453282H1SATMON028 g886470 BLASTN 608 1e−46 81 1255 456 700224308H1 SATMON011g1814402 BLASTN 661 1e−46 83 1256 456 700150484H1 SATMON007 g886470BLASTN 663 1e−46 78 1257 456 700240609H1 SATMON010 g2738247 BLASTN 6681e−46 79 1258 456 700171610H1 SATMON013 g974781 BLASTN 669 1e−46 79 1259456 LIB143-027-Q1-E1-H3 LIB143 g1814402 BLASTN 670 1e−46 86 1260 456700334084H1 SATMON019 g1814402 BLASTN 467 1e−45 81 1261 456LIB3069-043-Q1-K1-H6 LIB3069 g886470 BLASTN 546 1e−45 83 1262 456700222515H1 SATMON011 g1814402 BLASTN 648 1e−45 75 1263 456 700223749H1SATMON011 g1814402 BLASTN 648 1e−45 75 1264 456 701180187H1 SATMONN05g974781 BLASTN 648 1e−45 78 1265 456 700050111H1 SATMON003 g2738247BLASTN 653 1e−45 82 1266 456 700051275H1 SATMON003 g1814402 BLASTN 3791e−44 75 1267 456 700235202H1 SATMON010 g2738247 BLASTN 637 1e−44 801268 456 700257385H1 SATMON017 g1814402 BLASTN 638 1e−44 80 1269 456LIB143-028-Q1-E1-H7 LIB143 g1814402 BLASTN 638 1e−44 80 1270 456700576066H1 SATMON030 g886470 BLASTN 516 1e−43 76 1271 456 700455065H1SATMON029 g886470 BLASTN 624 1e−43 79 1272 456 700074158H1 SATMON007g1814402 BLASTN 630 1e−43 83 1273 456 700171322H1 SATMON013 g1814402BLASTN 634 1e−43 77 1274 456 700457369H1 SATMON029 g974781 BLASTN 3921e−42 83 1275 456 700378252H1 SATMON019 g2738247 BLASTN 462 1e−42 771276 456 700454626H1 SATMON029 g1814402 BLASTN 495 1e−42 85 1277 456700208182H1 SATMON016 g2738247 BLASTN 564 1e−42 73 1278 456 700618848H1SATMON034 g886470 BLASTN 615 1e−42 80 1279 456 700222969H1 SATMON011g1814402 BLASTN 622 1e−42 74 1280 456 700444782H1 SATMON027 g1814402BLASTN 337 1e−41 81 1281 456 700204404H1 SATMON003 g2738247 BLASTN 6071e−41 80 1282 456 700347028H1 SATMON021 g1814402 BLASTN 598 1e−40 811283 456 700172447H1 SATMON013 g1814402 BLASTN 328 1e−39 85 1284 456700549696H1 SATMON022 g886470 BLASTN 360 1e−39 81 1285 456 700257287H1SATMON017 g1814402 BLASTN 365 1e−38 84 1286 456 700569630H1 SATMON030g1814402 BLASTN 566 1e−38 78 1287 456 700207258H1 SATMON017 g1814402BLASTN 568 1e−38 85 1288 456 700052467H1 SATMON003 g1814402 BLASTN 5151e−37 82 1289 456 700083656H1 SATMON011 g2738247 BLASTN 552 1e−37 831290 456 700429279H1 SATMONN01 g2738247 BLASTN 556 1e−37 79 1291 456700349921H1 SATMON023 g2738247 BLASTN 471 1e−36 78 1292 456 700449609H1SATMON028 g886470 BLASTN 540 1e−36 81 1293 456 700150152H1 SATMON007g1814402 BLASTN 543 1e−36 74 1294 456 700075675H1 SATMON007 g1814402BLASTN 546 1e−36 81 1295 456 700267015H1 SATMON017 g886470 BLASTN 5501e−36 75 1296 456 700465118H1 SATMON025 g1814402 BLASTN 555 1e−36 781297 456 700456662H1 SATMON029 g2738247 BLASTN 314 1e−34 76 1298 456700405315H1 SATMON029 g886470 BLASTN 519 1e−34 83 1299 456 700218639H1SATMON011 g974781 BLASTN 524 1e−34 81 1300 456 700216606H1 SATMON016g1814402 BLASTN 524 1e−34 83 1301 456 700456965H1 SATMON029 g2738247BLASTN 525 1e−34 78 1302 456 700102147H1 SATMON010 g974781 BLASTN 5261e−34 82 1303 456 700235734H1 SATMON010 g2738247 BLASTN 526 1e−34 831304 456 LIB3068-009-Q1-K1-E10 LIB3068 g886470 BLASTN 548 1e−34 70 1305456 700029363H1 SATMON003 g974781 BLASTN 377 1e−33 80 1306 456LIB3069-030-Q1-K1-D10 LIB3069 g886470 BLASTN 297 1e−30 78 1307 456700334895H1 SATMON019 g974781 BLASTN 468 1e−30 82 1308 456 700150963H1SATMON007 g974781 BLASTN 459 1e−29 88 1309 456 700152817H1 SATMON007g974781 BLASTN 459 1e−29 88 1310 456 700208732H1 SATMON016 g974781BLASTN 485 1e−29 76 1311 456 700051461H1 SATMON003 g886470 BLASTN 4671e−28 73 1312 456 700616589H1 SATMON033 g2738247 BLASTN 273 1e−27 781313 456 700236150H1 SATMON010 g974781 BLASTN 434 1e−27 84 1314 456700453369H1 SATMON028 g1814402 BLASTN 436 1e−27 84 1315 456 700621739H1SATMON034 g1814402 BLASTN 436 1e−27 69 1316 456 700075162H1 SATMON007g1814402 BLASTN 438 1e−27 85 1317 456 700405040H1 SATMON027 g974781BLASTN 438 1e−27 72 1318 456 700356893H1 SATMON024 g974781 BLASTN 4581e−27 82 1319 456 701185493H1 SATMONN06 g886471 BLASTX 72 1e−26 97 1320456 700206095H1 SATMON003 g974781 BLASTN 421 1e−26 83 1321 456700083901H1 SATMON011 g1814402 BLASTN 421 1e−26 80 1322 456LIB3062-026-Q1-K1-D7 LIB3062 g1814402 BLASTN 423 1e−24 81 1323 456700377277H1 SATMON019 g1814402 BLASTN 357 1e−20 75 1324 456 700201214H1SATMON003 g2738248 BLASTX 152 1e−18 77 1325 456 700025959H1 SATMON003g886471 BLASTX 188 1e−18 94 1326 456 700259484H1 SATMON017 g1814402BLASTN 189 1e−17 82 1327 456 700613969H1 SATMON033 g886470 BLASTN 3151e−17 90 1328 456 700215602H1 SATMON016 g2738248 BLASTX 158 1e−16 741329 456 700438152H1 SATMON026 g2738247 BLASTN 258 1e−16 75 1330 456700458654H1 SATMON029 g886470 BLASTN 330 1e−16 70 1331 456 700449563H1SATMON028 g886471 BLASTX 163 1e−15 89 1332 456 700236490H1 SATMON010g1814402 BLASTN 276 1e−14 89 1333 456 700456927H1 SATMON029 g1814403BLASTX 119 1e−13 86 1334 456 701180088H1 SATMONN05 g886471 BLASTX 1521e−13 60 1335 456 700455716H1 SATMON029 g2738248 BLASTX 111 1e−12 671336 456 700573520H1 SATMON030 g886471 BLASTX 82 1e−11 84 1337 456700569938H1 SATMON030 g2738248 BLASTX 132 1e−11 91 1338 456 700551958H1SATMON022 g2738247 BLASTN 264 1e−11 82 1339 456 700337475H1 SATMON020g2738247 BLASTN 268 1e−11 83 1340 456 700170827H1 SATMON013 g886471BLASTX 124 1e−10 77 1341 456 700453382H1 SATMON028 g974781 BLASTN 1521e−10 85 1342 456 700089211H1 SATMON011 g2738247 BLASTN 245 1e−9 79 1343456 700103155H1 SATMON010 g2738248 BLASTX 82 1e−8 85 1344 5523LIB3062-017-Q1-K1-F4 LIB3062 g2738247 BLASTN 932 1e−84 76 1345 5523700210708H1 SATMON016 g2738247 BLASTN 952 1e−70 79 1346 5523 700219307H1SATMON011 g1814402 BLASTN 892 1e−65 78 1347 5523 700221188H1 SATMON011g2738247 BLASTN 867 1e−63 81 1348 5523 700203884H1 SATMON003 g2738247BLASTN 874 1e−63 77 1349 5523 700218549H1 SATMON011 g2738247 BLASTN 8111e−58 77 1350 5523 700572845H2 SATMON030 g2738247 BLASTN 785 1e−56 771351 5523 700152362H1 SATMON007 g886470 BLASTN 742 1e−52 79 1352 5523700152138H1 SATMON007 g2738247 BLASTN 714 1e−50 80 1353 5523 700218842H1SATMON011 g2738247 BLASTN 700 1e−49 75 2666 -700697958 700697958H1SOYMON015 g2738247 BLASTN 258 1e−10 83 2667 -700731277 700731277H1SOYMON009 g886470 BLASTN 930 1e−68 83 2668 -700749261 700749261H1SOYMON013 g1814402 BLASTN 508 1e−38 78 2669 -700787742 700787742H2SOYMON011 g1814402 BLASTN 648 1e−50 81 2670 -700831146 700831146H1SOYMON019 g1814402 BLASTN 476 1e−34 80 2671 -700832029 700832029H1SOYMON019 g1814402 BLASTN 328 1e−27 78 2672 -700854481 700854481H1SOYMON023 g1814402 BLASTN 193 1e−11 85 2673 -700873716 700873716H1SOYMON018 g1749542 BLASTX 99 1e−15 53 2674 -700893904 700893904H1SOYMON024 g886470 BLASTN 413 1e−25 82 2675 -700909658 700909658H1SOYMON022 g1814402 BLASTN 251 1e−10 91 2676 -700943841 700943841H1SOYMON024 g886470 BLASTN 549 1e−49 82 2677 -700963223 700963223H1SOYMON022 g886470 BLASTN 649 1e−45 72 2678 -700974103 700974103H1SOYMON005 g886470 BLASTN 763 1e−54 80 2679 -700994293 700994293H1SOYMON011 g974782 BLASTX 97 1e−12 72 2680 -701004816 701004816H1SOYMON019 g1814402 BLASTN 582 1e−39 84 2681 -701007125 701007125H1SOYMON019 g1814403 BLASTX 152 1e−13 87 2682 -701008540 701008540H1SOYMON019 g886470 BLASTN 770 1e−55 74 2683 -701037766 701037766H1SOYMON029 g1814403 BLASTX 72 1e−10 69 2684 -701062191 701062191H1SOYMON033 g974781 BLASTN 225 1e−9 79 2685 -701105474 701105474H1SOYMON036 g974782 BLASTX 164 1e−15 91 2686 -GM14442 LIB3049-056-Q1-E1-B1LIB3049 g974781 BLASTN 231 1e−10 79 2687 -GM19631 LIB3056-007-Q1-N1-G9LIB3056 g974781 BLASTN 388 1e−37 78 2688 -GM37189 LIB3051-072-Q1-K1-B5LIB3051 g1814402 BLASTN 316 1e−15 78 2689 -GM44802 LIB3053-004-Q1-N1-B2LIB3053 g974782 BLASTX 85 1e−26 90 2690 1382 700683236H1 SOYMON008g886470 BLASTN 805 1e−58 78 2691 1382 700566434H1 SOYMON002 g886471BLASTX 162 1e−23 76 2692 15690 701064361H1 SOYMON034 g974781 BLASTN 9511e−70 84 2693 15690 700847301H1 SOYMON021 g886470 BLASTN 707 1e−66 852694 17335 LIB3051-088-Q1-K1-D9 LIB3051 g886470 BLASTN 1285 1e−101 792695 17335 701003832H1 SOYMON019 g974781 BLASTN 811 1e−58 77 2696 17335700864888H1 SOYMON016 g1814402 BLASTN 767 1e−55 79 2697 17335700672491H1 SOYMON006 g974781 BLASTN 428 1e−46 75 2698 17335 701003457H1SOYMON019 g1814402 BLASTN 411 1e−29 84 2699 17335 700833672H1 SOYMON019g974781 BLASTN 445 1e−28 79 2700 17900 700850578H1 SOYMON023 g2738247BLASTN 905 1e−66 82 2701 17900 701053154H1 SOYMON032 g1814402 BLASTN 7251e−65 83 2702 17900 700842351H1 SOYMON020 g1814402 BLASTN 878 1e−64 832703 17900 700837656H1 SOYMON020 g1814402 BLASTN 841 1e−61 83 2704 17900700890851H1 SOYMON024 g974781 BLASTN 747 1e−53 80 2705 20688 700908840H1SOYMON022 g2738247 BLASTN 889 1e−65 82 2706 20688 700908848H1 SOYMON022g2738247 BLASTN 877 1e−64 81 2707 33542 LIB3051-009-Q1-E1-E5 LIB3051g974781 BLASTN 1218 1e−92 79 2708 33542 700748773H1 SOYMON013 g974781BLASTN 669 1e−46 78 2709 33542 700836363H1 SOYMON020 g2738247 BLASTN 5961e−40 80 2710 4243 701123616H1 SOYMON037 g1814402 BLASTN 995 1e−74 872711 4243 700555001H1 SOYMON001 g1814402 BLASTN 975 1e−72 90 2712 4243701002967H1 SOYMON019 g1814402 BLASTN 950 1e−70 86 2713 4243 700653509H1SOYMON003 g1814402 BLASTN 539 1e−69 85 2714 4243 701206028H1 SOYMON035g1814402 BLASTN 938 1e−69 86 2715 4243 700962115H1 SOYMON022 g1814402BLASTN 923 1e−68 86 2716 4243 700866243H1 SOYMON016 g1814402 BLASTN 9091e−66 82 2717 4243 700752507H1 SOYMON014 g1814402 BLASTN 887 1e−65 852718 4243 701003887H1 SOYMON019 g1814402 BLASTN 863 1e−63 86 2719 4243700556913H1 SOYMON001 g1814402 BLASTN 865 1e−63 86 2720 4243 701013549H1SOYMON019 g1814402 BLASTN 867 1e−63 90 2721 4243 701209706H1 SOYMON035g1814402 BLASTN 871 1e−63 90 2722 4243 701010487H1 SOYMON019 g1814402BLASTN 529 1e−62 79 2723 4243 700548246H1 SOYMON002 g1814402 BLASTN 5531e−62 82 2724 4243 701138219H1 SOYMON038 g1814402 BLASTN 594 1e−62 862725 4243 700965160H1 SOYMON022 g1814402 BLASTN 852 1e−62 91 2726 4243701015168H1 SOYMON019 g1814402 BLASTN 855 1e−62 89 2727 4243 701136095H1SOYMON038 g1814402 BLASTN 839 1e−61 88 2728 4243 700761789H1 SOYMON015g1814402 BLASTN 845 1e−61 86 2729 4243 701105695H1 SOYMON036 g1814402BLASTN 835 1e−60 87 2730 4243 700991714H1 SOYMON011 g1814402 BLASTN 5021e−59 83 2731 4243 700564223H1 SOYMON002 g1814402 BLASTN 557 1e−59 902732 4243 700987384H1 SOYMON009 g1814402 BLASTN 803 1e−58 86 2733 4243700833934H1 SOYMON019 g1814402 BLASTN 806 1e−58 89 2734 4243 700835181H1SOYMON019 g1814402 BLASTN 806 1e−58 82 2735 4243 700737529H1 SOYMON010g1814402 BLASTN 810 1e−58 92 2736 4243 701012851H1 SOYMON019 g1814402BLASTN 811 1e−58 91 2737 4243 700556592H1 SOYMON001 g1814402 BLASTN 8141e−58 88 2738 4243 700907579H1 SOYMON022 g1814402 BLASTN 781 1e−56 892739 4243 700961749H1 SOYMON022 g1814402 BLASTN 785 1e−56 91 2740 4243700835239H1 SOYMON019 g1814402 BLASTN 787 1e−56 86 2741 4243 700646425H1SOYMON013 g1814402 BLASTN 772 1e−55 89 2742 4243 701123924H1 SOYMON037g1814402 BLASTN 775 1e−55 91 2743 4243 700957759H1 SOYMON022 g1814402BLASTN 776 1e−55 90 2744 4243 700964425H1 SOYMON022 g1814402 BLASTN 7771e−55 86 2745 4243 700962173H1 SOYMON022 g1814402 BLASTN 778 1e−55 912746 4243 701066293H1 SOYMON034 g1814402 BLASTN 759 1e−54 81 2747 4243700986741H1 SOYMON009 g1814402 BLASTN 589 1e−53 84 2748 4243 701212514H1SOYMON035 g1814402 BLASTN 591 1e−53 87 2749 4243 701009195H1 SOYMON019g1814402 BLASTN 747 1e−53 91 2750 4243 701060510H1 SOYMON033 g1814402BLASTN 748 1e−53 84 2751 4243 700848730H1 SOYMON021 g1814402 BLASTN 7491e−53 90 2752 4243 700754870H1 SOYMON014 g1814402 BLASTN 751 1e−53 842753 4243 701212482H1 SOYMON035 g1814402 BLASTN 751 1e−53 84 2754 4243700753283H1 SOYMON014 g1814402 BLASTN 752 1e−53 86 2755 4243 700738517H1SOYMON012 g1814402 BLASTN 636 1e−52 89 2756 4243 700833978H1 SOYMON019g1814402 BLASTN 740 1e−52 91 2757 4243 700756424H1 SOYMON014 g1814402BLASTN 729 1e−51 82 2758 4243 701011759H1 SOYMON019 g1814402 BLASTN 7291e−51 91 2759 4243 701010103H2 SOYMON019 g1814402 BLASTN 707 1e−50 822760 4243 700741392H1 SOYMON012 g1814402 BLASTN 707 1e−50 84 2761 4243701123062H1 SOYMON037 g1814402 BLASTN 308 1e−49 88 2762 4243 701048949H1SOYMON032 g1814402 BLASTN 502 1e−49 85 2763 4243 700834566H1 SOYMON019g1814402 BLASTN 618 1e−49 88 2764 4243 700963965H1 SOYMON022 g1814402BLASTN 685 1e−48 78 2765 4243 700986376H1 SOYMON009 g1814402 BLASTN 6941e−48 84 2766 4243 701012708H1 SOYMON019 g1814402 BLASTN 521 1e−47 912767 4243 700746927H1 SOYMON013 g1814402 BLASTN 547 1e−46 78 2768 4243700997448H1 SOYMON018 g1814402 BLASTN 470 1e−45 89 2769 4243 700830667H1SOYMON019 g1814402 BLASTN 647 1e−45 86 2770 4243 700891479H1 SOYMON024g1814402 BLASTN 654 1e−45 88 2771 4243 700562246H1 SOYMON002 g1814402BLASTN 656 1e−45 86 2772 4243 701097325H1 SOYMON028 g1814402 BLASTN 4201e−44 92 2773 4243 700835830H1 SOYMON019 g1814402 BLASTN 503 1e−44 862774 4243 700962969H1 SOYMON022 g1814402 BLASTN 515 1e−44 89 2775 4243701102860H1 SOYMON028 g1814402 BLASTN 309 1e−42 92 2776 4243 700763506H1SOYMON015 g1814402 BLASTN 456 1e−42 84 2777 4243 700994059H1 SOYMON011g1814402 BLASTN 487 1e−42 87 2778 4243 700751551H1 SOYMON014 g1814402BLASTN 568 1e−41 85 2779 4243 701108872H1 SOYMON036 g1814402 BLASTN 6031e−41 85 2780 4243 700994053H1 SOYMON011 g1814402 BLASTN 587 1e−40 852781 4243 700729064H1 SOYMON009 g1814402 BLASTN 413 1e−38 90 2782 4243700874176H1 SOYMON018 g1814402 BLASTN 468 1e−38 92 2783 4243 700742963H1SOYMON012 g1814402 BLASTN 565 1e−38 86 2784 4243 701212923H1 SOYMON035g1814402 BLASTN 572 1e−38 84 2785 4243 700994851H1 SOYMON011 g1814402BLASTN 548 1e−36 76 2786 4243 701000193H1 SOYMON018 g1814402 BLASTN 2961e−33 87 2787 4243 700756219H1 SOYMON014 g1814402 BLASTN 394 1e−32 802788 4243 701014751H1 SOYMON019 g1814402 BLASTN 461 1e−29 90 2789 4243700561163H1 SOYMON002 g1814402 BLASTN 231 1e−25 89 2790 4243 700650284H1SOYMON003 g974782 BLASTX 157 1e−14 96 2791 4243 700869218H1 SOYMON016g1814402 BLASTN 248 1e−11 93 2792 550 LIB3028-007-Q1-B1-B6 LIB3028g886470 BLASTN 1440 1e−111 81 2793 550 LIB3040-017-Q1-E1-E8 LIB3040g886470 BLASTN 1260 1e−96 80 2794 550 700650656H1 SOYMON003 g1814402BLASTN 1072 1e−93 83 2795 550 LIB3051-091-Q1-K1-C2 LIB3051 g886470BLASTN 1124 1e−89 80 2796 550 LIB3051-072-Q1-K1-B3 LIB3051 g974781BLASTN 1114 1e−83 82 2797 550 LIB3051-006-Q1-E1-G9 LIB3051 g974781BLASTN 703 1e−82 81 2798 550 700563811H1 SOYMON002 g1814402 BLASTN 10711e−80 84 2799 550 700754104H1 SOYMON014 g974781 BLASTN 1052 1e−78 862800 550 701002973H1 SOYMON019 g1814402 BLASTN 1039 1e−77 85 2801 550700986733H1 SOYMON009 g974781 BLASTN 1039 1e−77 83 2802 550 700557595H1SOYMON001 g886470 BLASTN 1019 1e−76 83 2803 550 700563477H1 SOYMON002g2738247 BLASTN 1024 1e−76 85 2804 550 700976112H1 SOYMON009 g886470BLASTN 1024 1e−76 84 2805 550 701004484H1 SOYMON019 g1814402 BLASTN 10071e−75 84 2806 550 700889161H1 SOYMON024 g974781 BLASTN 996 1e−74 85 2807550 700833110H1 SOYMON019 g974781 BLASTN 998 1e−74 87 2808 550701124559H1 SOYMON037 g974781 BLASTN 1002 1e−74 86 2809 550 700729207H1SOYMON009 g974781 BLASTN 1004 1e−74 85 2810 550 700730012H1 SOYMON009g886470 BLASTN 987 1e−73 85 2811 550 700987128H1 SOYMON009 g2738247BLASTN 989 1e−73 85 2812 550 700564788H1 SOYMON002 g974781 BLASTN 9901e−73 84 2813 550 701099104H1 SOYMON028 g1814402 BLASTN 994 1e−73 862814 550 LIB3065-008-Q1-N1-B4 LIB3065 g2738247 BLASTN 839 1e−72 83 2815550 701104283H1 SOYMON036 g974781 BLASTN 976 1e−72 83 2816 550700726387H1 SOYMON009 g974781 BLASTN 980 1e−72 84 2817 550 700900884H1SOYMON027 g1814402 BLASTN 982 1e−72 85 2818 550 LIB3051-029-Q1-K1-D6LIB3051 g886470 BLASTN 824 1e−71 81 2819 550 701213995H1 SOYMON035g1814402 BLASTN 962 1e−71 84 2820 550 700683596H1 SOYMON008 g1814402BLASTN 968 1e−71 85 2821 550 700646529H1 SOYMON014 g1814402 BLASTN 8671e−70 84 2822 550 700756704H1 SOYMON014 g1814402 BLASTN 893 1e−70 872823 550 700991647H1 SOYMON011 g1814402 BLASTN 947 1e−70 84 2824 550700962195H1 SOYMON022 g974781 BLASTN 947 1e−70 87 2825 550 700994369H1SOYMON011 g1814402 BLASTN 949 1e−70 84 2826 550 700672477H1 SOYMON006g1814402 BLASTN 951 1e−70 85 2827 550 700946215H1 SOYMON024 g2738247BLASTN 957 1e−70 83 2828 550 701152765H1 SOYMON031 g974781 BLASTN 9581e−70 88 2829 550 701010552H1 SOYMON019 g974781 BLASTN 501 1e−69 83 2830550 LIB3056-004-Q1-N1-B12 LIB3056 g2738247 BLASTN 698 1e−69 77 2831 550701100656H1 SOYMON028 g974781 BLASTN 890 1e−69 85 2832 550 700985362H1SOYMON009 g1814402 BLASTN 937 1e−69 82 2833 550 700746178H1 SOYMON013g974781 BLASTN 938 1e−69 85 2834 550 700674440H1 SOYMON007 g974781BLASTN 938 1e−69 83 2835 550 700556934H1 SOYMON001 g1814402 BLASTN 9381e−69 82 2836 550 700652624H1 SOYMON003 g1814402 BLASTN 939 1e−69 822837 550 700952331H1 SOYMON022 g886470 BLASTN 946 1e−69 83 2838 550700725576H1 SOYMON009 g886470 BLASTN 946 1e−69 85 2839 550 701003602H1SOYMON019 g2738247 BLASTN 924 1e−68 83 2840 550 700745053H1 SOYMON013g974781 BLASTN 924 1e−68 85 2841 550 700895781H1 SOYMON027 g1814402BLASTN 926 1e−68 84 2842 550 700664593H1 SOYMON005 g974781 BLASTN 9261e−68 87 2843 550 700864264H1 SOYMON016 g1814402 BLASTN 930 1e−68 842844 550 700674466H1 SOYMON007 g974781 BLASTN 933 1e−68 83 2845 550700564170H1 SOYMON002 g974781 BLASTN 861 1e−67 83 2846 550 700654531H1SOYMON004 g974781 BLASTN 911 1e−67 81 2847 550 700996341H1 SOYMON018g886470 BLASTN 911 1e−67 83 2848 550 701136257H1 SOYMON038 g886470BLASTN 912 1e−67 82 2849 550 700751114H1 SOYMON014 g1814402 BLASTN 9141e−67 84 2850 550 700657606H1 SOYMON004 g1814402 BLASTN 916 1e−67 872851 550 700983827H1 SOYMON009 g974781 BLASTN 918 1e−67 85 2852 550700981249H1 SOYMON009 g1814402 BLASTN 921 1e−67 80 2853 550 700836103H1SOYMON019 g886470 BLASTN 922 1e−67 82 2854 550 701011869H1 SOYMON019g886470 BLASTN 486 1e−66 86 2855 550 701097045H1 SOYMON028 g886470BLASTN 601 1e−66 85 2856 550 701012695H1 SOYMON019 g974781 BLASTN 7771e−66 85 2857 550 700945396H1 SOYMON024 g974781 BLASTN 902 1e−66 85 2858550 700755390H1 SOYMON014 g974781 BLASTN 903 1e−66 86 2859 550700967721H1 SOYMON033 g886470 BLASTN 910 1e−66 82 2860 550 700750610H1SOYMON014 g974781 BLASTN 910 1e−66 84 2861 550 700908231H1 SOYMON022g974781 BLASTN 527 1e−65 83 2862 550 701003835H1 SOYMON019 g886470BLASTN 723 1e−65 82 2863 550 700790802H1 SOYMON011 g2738247 BLASTN 8911e−65 82 2864 550 701053438H1 SOYMON032 g974781 BLASTN 893 1e−65 82 2865550 701009430H1 SOYMON019 g974781 BLASTN 894 1e−65 85 2866 550700891415H1 SOYMON024 g1814402 BLASTN 895 1e−65 84 2867 550 701100681H1SOYMON028 g2738247 BLASTN 482 1e−64 84 2868 550 700893420H1 SOYMON024g974781 BLASTN 570 1e−64 89 2869 550 700752741H1 SOYMON014 g886470BLASTN 880 1e−64 82 2870 550 700955938H1 SOYMON022 g1814402 BLASTN 8821e−64 81 2871 550 701015042H1 SOYMON019 g2738247 BLASTN 886 1e−64 822872 550 LIB3051-072-Q1-K1-B1 LIB3051 g974781 BLASTN 645 1e−63 76 2873550 700741918H1 SOYMON012 g886470 BLASTN 722 1e−63 84 2874 550701103319H1 SOYMON028 g974781 BLASTN 792 1e−63 82 2875 550 700833218H1SOYMON019 g2738247 BLASTN 863 1e−63 81 2876 550 701008071H1 SOYMON019g886470 BLASTN 867 1e−63 83 2877 550 700832073H1 SOYMON019 g974781BLASTN 871 1e−63 83 2878 550 700889695H1 SOYMON024 g974781 BLASTN 8721e−63 83 2879 550 701007489H2 SOYMON019 g974781 BLASTN 873 1e−63 84 2880550 700895858H1 SOYMON027 g974781 BLASTN 874 1e−63 84 2881 550700753955H1 SOYMON014 g974781 BLASTN 470 1e−62 87 2882 550 700564433H1SOYMON002 g886470 BLASTN 757 1e−62 84 2883 550 700963115H1 SOYMON022g974781 BLASTN 851 1e−62 83 2884 550 700894728H1 SOYMON024 g886470BLASTN 860 1e−62 84 2885 550 701056915H1 SOYMON033 g886470 BLASTN 8621e−62 84 2886 550 700741134H1 SOYMON012 g886470 BLASTN 862 1e−62 83 2887550 700847591H1 SOYMON021 g974781 BLASTN 842 1e−61 84 2888 550700941253H1 SOYMON024 g2738247 BLASTN 844 1e−61 80 2889 550 701004315H1SOYMON019 g2738247 BLASTN 846 1e−61 83 2890 550 700895720H1 SOYMON027g974781 BLASTN 848 1e−61 83 2891 550 701013541H1 SOYMON019 g974781BLASTN 849 1e−61 84 2892 550 700892552H1 SOYMON024 g886470 BLASTN 7191e−60 83 2893 550 701141313H1 SOYMON038 g974781 BLASTN 827 1e−60 83 2894550 701012547H1 SOYMON019 g1814402 BLASTN 831 1e−60 83 2895 550701008558H1 SOYMON019 g1814402 BLASTN 831 1e−60 83 2896 550 700902022H1SOYMON027 g2738247 BLASTN 831 1e−60 82 2897 550 700959515H1 SOYMON022g886470 BLASTN 832 1e−60 81 2898 550 701042630H1 SOYMON029 g1814402BLASTN 819 1e−59 81 2899 550 700941292H1 SOYMON024 g2738247 BLASTN 8201e−59 81 2900 550 700788526H1 SOYMON011 g974781 BLASTN 491 1e−58 82 2901550 700894839H1 SOYMON024 g1814402 BLASTN 498 1e−58 82 2902 550700865873H1 SOYMON016 g974781 BLASTN 808 1e−58 85 2903 550 701015435H1SOYMON019 g886470 BLASTN 809 1e−58 80 2904 550 700755960H1 SOYMON014g2738247 BLASTN 809 1e−58 78 2905 550 700876051H1 SOYMON018 g886470BLASTN 434 1e−57 81 2906 550 701041327H1 SOYMON029 g886470 BLASTN 4991e−57 83 2907 550 701098902H1 SOYMON028 g2738247 BLASTN 767 1e−57 772908 550 700853392H1 SOYMON023 g886470 BLASTN 793 1e−57 82 2909 550700872645H1 SOYMON018 g974781 BLASTN 798 1e−57 83 2910 550 700989675H1SOYMON011 g2738247 BLASTN 800 1e−57 79 2911 550 700753487H1 SOYMON014g1814402 BLASTN 589 1e−56 83 2912 550 700736276H1 SOYMON010 g1814402BLASTN 782 1e−56 79 2913 550 700891361H1 SOYMON024 g1814402 BLASTN 7831e−56 81 2914 550 700829712H1 SOYMON019 g974781 BLASTN 790 1e−56 80 2915550 LIB3050-019-Q1-K1-A1 LIB3050 g974781 BLASTN 663 1e−55 81 2916 550701001013H1 SOYMON018 g1814402 BLASTN 768 1e−55 84 2917 550LIB3028-031-Q1-B1-G12 LIB3028 g886470 BLASTN 775 1e−55 82 2918 550701212782H1 SOYMON035 g886470 BLASTN 621 1e−54 82 2919 550 701008695H1SOYMON019 g974781 BLASTN 718 1e−54 80 2920 550 700990972H1 SOYMON011g1814402 BLASTN 766 1e−54 81 2921 550 700789576H2 SOYMON011 g886470BLASTN 766 1e−54 80 2922 550 700994266H1 SOYMON011 g1814402 BLASTN 4081e−53 85 2923 550 700731985H1 SOYMON010 g974781 BLASTN 590 1e−53 81 2924550 700907927H1 SOYMON022 g886470 BLASTN 612 1e−53 80 2925 550701012079H1 SOYMON019 g974781 BLASTN 669 1e−53 86 2926 550 700753939H1SOYMON014 g886470 BLASTN 690 1e−53 78 2927 550 701000754H1 SOYMON018g974781 BLASTN 745 1e−53 76 2928 550 701040287H1 SOYMON029 g886470BLASTN 747 1e−53 82 2929 550 700891329H1 SOYMON024 g974781 BLASTN 7491e−53 79 2930 550 700897258H1 SOYMON027 g886470 BLASTN 752 1e−53 86 2931550 700944949H1 SOYMON024 g974781 BLASTN 426 1e−52 82 2932 550701108671H1 SOYMON036 g1814402 BLASTN 731 1e−52 77 2933 550 700905233H1SOYMON022 g886470 BLASTN 732 1e−52 77 2934 550 700958589H1 SOYMON022g886470 BLASTN 738 1e−52 80 2935 550 700666436H1 SOYMON005 g2738247BLASTN 739 1e−52 82 2936 550 700829902H1 SOYMON019 g886470 BLASTN 7401e−52 82 2937 550 700740110H1 SOYMON012 g974781 BLASTN 742 1e−52 83 2938550 700989055H1 SOYMON011 g886470 BLASTN 538 1e−50 79 2939 550701213370H1 SOYMON035 g974781 BLASTN 599 1e−50 83 2940 550 701098072H1SOYMON028 g974781 BLASTN 709 1e−50 74 2941 550 700896128H1 SOYMON027g886470 BLASTN 714 1e−50 83 2942 550 701060755H1 SOYMON033 g974781BLASTN 716 1e−50 74 2943 550 700953594H1 SOYMON022 g1814402 BLASTN 6951e−49 78 2944 550 701046911H1 SOYMON032 g2738247 BLASTN 687 1e−48 812945 550 701065707H1 SOYMON034 g1814402 BLASTN 443 1e−47 84 2946 550700962114H1 SOYMON022 g886470 BLASTN 678 1e−47 84 2947 550 700831826H1SOYMON019 g2738247 BLASTN 682 1e−47 77 2948 550 701054296H1 SOYMON032g886470 BLASTN 454 1e−46 83 2949 550 700888738H1 SOYMON024 g974781BLASTN 552 1e−46 77 2950 550 700892022H1 SOYMON024 g886470 BLASTN 6051e−46 81 2951 550 700890275H1 SOYMON024 g2738247 BLASTN 666 1e−46 772952 550 LIB3051-006-Q1-K1-G9 LIB3051 g974781 BLASTN 670 1e−45 80 2953550 700889113H1 SOYMON024 g886470 BLASTN 582 1e−43 81 2954 550700952720H1 SOYMON022 g886470 BLASTN 611 1e−42 76 2955 550 701014761H1SOYMON019 g886470 BLASTN 318 1e−41 84 2956 550 700753882H1 SOYMON014g886470 BLASTN 381 1e−41 79 2957 550 700743792H1 SOYMON012 g1814402BLASTN 610 1e−41 86 2958 550 700990963H1 SOYMON011 g886470 BLASTN 5781e−39 77 2959 550 700941880H1 SOYMON024 g2738247 BLASTN 583 1e−39 812960 550 700898962H1 SOYMON027 g2738247 BLASTN 571 1e−38 80 2961 550700990865H1 SOYMON011 g2738247 BLASTN 467 1e−37 77 2962 550 700565779H1SOYMON002 g886470 BLASTN 557 1e−37 70 2963 550 700993903H1 SOYMON011g974781 BLASTN 562 1e−37 84 2964 550 700941589H1 SOYMON024 g2738247BLASTN 550 1e−36 81 2965 550 701052554H1 SOYMON032 g886470 BLASTN 5341e−35 67 2966 550 700991055H1 SOYMON011 g886470 BLASTN 356 1e−34 79 2967550 701010438H1 SOYMON019 g2738247 BLASTN 508 1e−33 79 2968 550700756634H1 SOYMON014 g2738247 BLASTN 494 1e−32 76 2969 550 701042980H1SOYMON029 g886470 BLASTN 461 1e−29 83 2970 550 700682940H1 SOYMON008g2738247 BLASTN 466 1e−29 83 2971 550 701049575H1 SOYMON032 g886470BLASTN 433 1e−27 82 2972 550 700982552H1 SOYMON009 g886470 BLASTN 3561e−25 78 2973 550 700675637H1 SOYMON007 g886470 BLASTN 375 1e−25 79 2974550 701142153H1 SOYMON038 g886470 BLASTN 377 1e−22 76 2975 550700682724H1 SOYMON008 g974781 BLASTN 361 1e−19 88 2976 550 701051764H1SOYMON032 g1814402 BLASTN 211 1e−17 80 2977 550 700867241H1 SOYMON016g2738248 BLASTX 152 1e−13 88 2978 550 701054954H1 SOYMON032 g2738248BLASTX 138 1e−12 86 2979 550 700790450H2 SOYMON011 g974781 BLASTN 2381e−10 80 2980 550 700653979H1 SOYMON003 g1814403 BLASTX 118 1e−9 92 2981550 700894218H1 SOYMON024 g2738248 BLASTX 122 1e−9 78 2982 550700863078H1 SOYMON022 g2738247 BLASTN 236 1e−8 78 2983 5758 701209304H1SOYMON035 g886470 BLASTN 766 1e−54 84 2984 5758 701106455H1 SOYMON036g886470 BLASTN 723 1e−51 82 2985 5758 700833538H1 SOYMON019 g1814402BLASTN 609 1e−43 83 2986 5758 701051425H1 SOYMON032 g886470 BLASTN 6291e−43 83 2987 5758 700654506H1 SOYMON004 g886470 BLASTN 438 1e−26 702988 5758 701047795H1 SOYMON032 g974782 BLASTX 161 1e−15 100 2989 5758701202409H1 SOYMON035 g1814402 BLASTN 310 1e−15 79 2990 8266 700558628H1SOYMON001 g886470 BLASTN 780 1e−56 74 2991 8266 701207720H1 SOYMON035g1814402 BLASTN 766 1e−54 74 2992 8266 700557429H1 SOYMON001 g1814402BLASTN 728 1e−51 75

ADENOSYLHOMOCYSTEINASE (EC 3.3.1.1) Seq No. Cluster ID CloneID LibraryNCBI gi Method Score P-value % Ident 1354 -700154280 700154280H1SATMON007 g170772 BLASTN 229 1e−23 77 1355 -L30594291LIB3059-032-Q1-K1-A8 LIB3059 g170772 BLASTN 516 1e−74 73 1356 -L30664307LIB3066-047-Q1-K1-E7 LIB3066 g170772 BLASTN 628 1e−49 70 1357 503LIB3079-013-Q1-K1-D3 LIB3079 g170772 BLASTN 1505 1e−139 89 1358 503LIB3062-017-Q1-K1-A10 LIB3062 g170772 BLASTN 1571 1e−129 89 1359 503LIB3067-001-Q1-K1-D11 LIB3067 g170772 BLASTN 1654 1e−129 89 1360 503LIB148-060-Q1-E1-B9 LIB148 g170772 BLASTN 1620 1e−126 90 1361 503LIB3069-043-Q1-K1-G4 LIB3069 g170772 BLASTN 1360 1e−124 84 1362 503LIB143-006-Q1-E1-F3 LIB143 g170772 BLASTN 831 1e−120 88 1363 503LIB189-009-Q1-E1-E9 LIB189 g170772 BLASTN 1551 1e−120 90 1364 503LIB3060-003-Q1-K1-F9 LIB3060 g170772 BLASTN 1538 1e−119 91 1365 503700089023H1 SATMON011 g170772 BLASTN 1495 1e−115 91 1366 503LIB143-061-Q1-E1-C9 LIB143 g170772 BLASTN 1474 1e−114 90 1367 503LIB3069-034-Q1-K1-E5 LIB3069 g170772 BLASTN 1483 1e−114 88 1368 503LIB143-061-Q1-E1-E5 LIB143 g170772 BLASTN 1440 1e−113 89 1369 503LIB3067-040-Q1-K1-H5 LIB3067 g170772 BLASTN 1467 1e−113 88 1370 503LIB3067-048-Q1-K1-A11 LIB3067 g170772 BLASTN 1207 1e−109 86 1371 503LIB3068-050-Q1-K1-G6 LIB3068 g170772 BLASTN 1279 1e−109 86 1372 503LIB3069-036-Q1-K1-F6 LIB3069 g170772 BLASTN 1310 1e−109 92 1373 503LIB3066-047-Q1-K1-H5 LIB3066 g170772 BLASTN 1353 1e−108 86 1374 503LIB3059-011-Q1-K1-A5 LIB3059 g170772 BLASTN 1401 1e−107 90 1375 503LIB3067-056-Q1-K1-E11 LIB3067 g170772 BLASTN 957 1e−106 86 1376 503LIB189-010-Q1-E1-E12 LIB189 g170772 BLASTN 1144 1e−106 88 1377 503700084426H1 SATMON011 g170772 BLASTN 1368 1e−105 91 1378 503 700086273H1SATMON011 g170772 BLASTN 1364 1e−104 91 1379 503 700573027H1 SATMON030g170772 BLASTN 939 1e−103 89 1380 503 700209360H1 SATMON016 g170772BLASTN 1350 1e−103 89 1381 503 700619916H1 SATMON034 g170772 BLASTN 10501e−102 89 1382 503 700086051H1 SATMON011 g170772 BLASTN 1336 1e−102 901383 503 LIB3069-034-Q1-K1-C8 LIB3069 g170772 BLASTN 1322 1e−101 86 1384503 700026324H1 SATMON003 g170772 BLASTN 1328 1e−101 91 1385 503700104549H1 SATMON010 g170772 BLASTN 836 1e−100 88 1386 503 700622108H1SATMON034 g170772 BLASTN 1158 1e−100 88 1387 503 700093980H1 SATMON008g170772 BLASTN 1312 1e−100 90 1388 503 700077427H1 SATMON007 g170772BLASTN 1317 1e−100 90 1389 503 700095389H1 SATMON008 g170772 BLASTN 13021e−99 91 1390 503 LIB3067-055-Q1-K1-D3 LIB3067 g170772 BLASTN 762 1e−9887 1391 503 LIB3060-054-Q1-K1-F6 LIB3060 g170772 BLASTN 1009 1e−98 831392 503 700083339H1 SATMON011 g170772 BLASTN 1282 1e−98 91 1393 503700102631H1 SATMON010 g170772 BLASTN 1283 1e−98 89 1394 503 700265625H1SATMON017 g170772 BLASTN 1286 1e−98 90 1395 503 700095002H1 SATMON008g170772 BLASTN 1289 1e−98 88 1396 503 700094761H1 SATMON008 g170772BLASTN 1289 1e−98 88 1397 503 700073832H1 SATMON007 g170772 BLASTN 12701e−97 88 1398 503 700047817H1 SATMON003 g170772 BLASTN 1272 1e−97 911399 503 700091149H1 SATMON011 g170772 BLASTN 1262 1e−96 89 1400 503700098584H1 SATMON009 g170772 BLASTN 1264 1e−96 91 1401 503 700085932H1SATMON011 g170772 BLASTN 1266 1e−96 89 1402 503 700094081H1 SATMON008g170772 BLASTN 1269 1e−96 91 1403 503 700202442H1 SATMON003 g170772BLASTN 1145 1e−95 88 1404 503 700049770H1 SATMON003 g170772 BLASTN 12461e−95 89 1405 503 LIB3079-020-Q1-K1-B12 LIB3079 g170772 BLASTN 12491e−95 85 1406 503 700090226H1 SATMON011 g170772 BLASTN 1252 1e−95 901407 503 700088191H1 SATMON011 g170772 BLASTN 1253 1e−95 90 1408 503700076743H1 SATMON007 g170772 BLASTN 1256 1e−95 90 1409 503LIB189-009-Q1-E1-E10 LIB189 g170772 BLASTN 789 1e−94 88 1410 503700072038H1 SATMON007 g170772 BLASTN 1237 1e−94 89 1411 503 700082967H1SATMON011 g170772 BLASTN 1239 1e−94 89 1412 503 LIB3068-062-Q1-K1-A3LIB3068 g170772 BLASTN 1148 1e−93 82 1413 503 700094713H1 SATMON008g170772 BLASTN 1222 1e−93 88 1414 503 700095620H1 SATMON008 g170772BLASTN 1173 1e−92 89 1415 503 700242509H1 SATMON010 g170772 BLASTN 12131e−92 92 1416 503 700071923H1 SATMON007 g170772 BLASTN 1214 1e−92 901417 503 700575314H1 SATMON030 g170772 BLASTN 1149 1e−91 88 1418 503700086654H1 SATMON011 g170772 BLASTN 1199 1e−91 90 1419 503 700241072H1SATMON010 g170772 BLASTN 1205 1e−91 91 1420 503 700047361H1 SATMON003g170772 BLASTN 1110 1e−90 89 1421 503 700217056H1 SATMON016 g170772BLASTN 1187 1e−90 91 1422 503 LIB3067-005-Q1-K1-F2 LIB3067 g170772BLASTN 1188 1e−90 84 1423 503 700075495H1 SATMON007 g170772 BLASTN 11931e−90 86 1424 503 700084783H1 SATMON011 g170772 BLASTN 1194 1e−90 911425 503 LIB143-059-Q1-E1-C3 LIB143 g170772 BLASTN 952 1e−89 84 1426 503700448818H1 SATMON028 g170772 BLASTN 1174 1e−89 91 1427 503 700159079H1SATMON012 g170772 BLASTN 1177 1e−89 92 1428 503 700094408H1 SATMON008g170772 BLASTN 1177 1e−89 92 1429 503 700348440H1 SATMON023 g170772BLASTN 1182 1e−89 89 1430 503 700077082H1 SATMON007 g170772 BLASTN 11821e−89 92 1431 503 700082111H1 SATMON011 g170772 BLASTN 1183 1e−89 891432 503 700239752H1 SATMON010 g170772 BLASTN 1164 1e−88 89 1433 503700029456H1 SATMON003 g170772 BLASTN 1166 1e−88 90 1434 503 700209430H1SATMON016 g170772 BLASTN 1170 1e−88 90 1435 503 700208660H1 SATMON016g170772 BLASTN 930 1e−87 90 1436 503 700213138H1 SATMON016 g170772BLASTN 1042 1e−87 89 1437 503 700102234H1 SATMON010 g170772 BLASTN 6341e−85 90 1438 503 700450479H1 SATMON028 g170772 BLASTN 1015 1e−85 901439 503 700095372H1 SATMON008 g170772 BLASTN 1126 1e−85 90 1440 503700218734H1 SATMON011 g170772 BLASTN 1128 1e−85 94 1441 503 700256858H1SATMON017 g170772 BLASTN 1133 1e−85 90 1442 503 700221063H1 SATMON011g170772 BLASTN 1137 1e−85 90 1443 503 700105439H1 SATMON010 g170772BLASTN 1118 1e−84 89 1444 503 700085174H1 SATMON011 g170772 BLASTN 11251e−84 90 1445 503 700242421H1 SATMON010 g170772 BLASTN 1125 1e−84 921446 503 700077492H1 SATMON007 g170772 BLASTN 774 1e−83 88 1447 503700209295H1 SATMON016 g170772 BLASTN 914 1e−83 89 1448 503 700455826H1SATMON029 g170772 BLASTN 978 1e−83 91 1449 503 700213370H1 SATMON016g170772 BLASTN 1110 1e−83 92 1450 503 700352095H1 SATMON023 g170772BLASTN 1111 1e−83 89 1451 503 700076842H1 SATMON007 g170772 BLASTN 11121e−83 90 1452 503 700215872H1 SATMON016 g170772 BLASTN 1113 1e−83 891453 503 700073645H1 SATMON007 g170772 BLASTN 1113 1e−83 89 1454 503700048153H1 SATMON003 g170772 BLASTN 651 1e−82 90 1455 503 700073307H1SATMON007 g170772 BLASTN 762 1e−82 90 1456 503 700350009H1 SATMON023g170772 BLASTN 924 1e−82 88 1457 503 LIB143-059-Q1-E1-C5 LIB143 g170772BLASTN 1029 1e−82 83 1458 503 700073955H1 SATMON007 g170772 BLASTN 10901e−82 90 1459 503 LIB3059-042-Q1-K1-H12 LIB3059 g170772 BLASTN 10901e−82 85 1460 503 700238024H1 SATMON010 g170772 BLASTN 1091 1e−82 901461 503 700155863H1 SATMON007 g170772 BLASTN 1091 1e−82 93 1462 503700217890H1 SATMON016 g170772 BLASTN 1093 1e−82 91 1463 503 700239314H1SATMON010 g170772 BLASTN 1094 1e−82 88 1464 503 700048337H1 SATMON003g170772 BLASTN 1094 1e−82 92 1465 503 700235469H1 SATMON010 g170772BLASTN 1098 1e−82 91 1466 503 700164224H1 SATMON013 g170772 BLASTN 7101e−81 92 1467 503 700209374H1 SATMON016 g170772 BLASTN 1083 1e−81 891468 503 700082268H1 SATMON011 g170772 BLASTN 1083 1e−81 88 1469 503700159326H1 SATMON012 g170772 BLASTN 777 1e−80 91 1470 503 700025934H1SATMON003 g170772 BLASTN 792 1e−80 91 1471 503 700210458H1 SATMON016g170772 BLASTN 856 1e−80 88 1472 503 700243826H1 SATMON010 g170772BLASTN 881 1e−80 90 1473 503 700242676H1 SATMON010 g170772 BLASTN 9401e−80 89 1474 503 700345565H1 SATMON021 g170772 BLASTN 1034 1e−80 891475 503 700264733H1 SATMON017 g170772 BLASTN 1044 1e−80 90 1476 503700159161H1 SATMON012 g170772 BLASTN 1068 1e−80 90 1477 503 700072491H1SATMON007 g170772 BLASTN 604 1e−79 88 1478 503 700053204H1 SATMON008g170772 BLASTN 618 1e−79 89 1479 503 700451515H1 SATMON028 g170772BLASTN 1057 1e−79 85 1480 503 700468929H1 SATMON025 g170772 BLASTN 10611e−79 84 1481 503 700205801H1 SATMON003 g170772 BLASTN 1064 1e−79 911482 503 700611326H1 SATMON022 g170772 BLASTN 725 1e−78 88 1483 503700082291H1 SATMON011 g170772 BLASTN 1043 1e−78 87 1484 503 700071695H1SATMON007 g170772 BLASTN 1048 1e−78 89 1485 503 700551561H1 SATMON022g170772 BLASTN 746 1e−77 88 1486 503 700221019H1 SATMON011 g170772BLASTN 856 1e−77 89 1487 503 LIB3078-007-Q1-K1-C5 LIB3078 g170772 BLASTN936 1e−77 83 1488 503 700049925H1 SATMON003 g170772 BLASTN 954 1e−77 891489 503 700154101H1 SATMON007 g170772 BLASTN 1036 1e−77 88 1490 503700380151H1 SATMON021 g170772 BLASTN 613 1e−76 88 1491 503 700243831H1SATMON010 g170772 BLASTN 842 1e−76 89 1492 503 700235671H1 SATMON010g170772 BLASTN 1019 1e−76 90 1493 503 700087847H1 SATMON011 g170772BLASTN 1009 1e−75 82 1494 503 700208747H1 SATMON016 g170772 BLASTN 10101e−75 89 1495 503 700071795H1 SATMON007 g170772 BLASTN 1011 1e−75 891496 503 700157002H1 SATMON012 g170772 BLASTN 1012 1e−75 90 1497 503700201608H1 SATMON003 g170772 BLASTN 1015 1e−75 85 1498 503 700451541H1SATMON028 g170772 BLASTN 1015 1e−75 86 1499 503 700212182H1 SATMON016g170772 BLASTN 1015 1e−75 88 1500 503 700381485H1 SATMON023 g170772BLASTN 1017 1e−75 91 1501 503 700096793H1 SATMON008 g170772 BLASTN 7611e−74 87 1502 503 700093025H1 SATMON008 g170772 BLASTN 996 1e−74 89 1503503 700017129H1 SATMON001 g170772 BLASTN 998 1e−74 92 1504 503700216525H1 SATMON016 g170772 BLASTN 1000 1e−74 86 1505 503 700087912H1SATMON011 g170772 BLASTN 1001 1e−74 91 1506 503 700172618H1 SATMON013g170772 BLASTN 1001 1e−74 91 1507 503 700801894H1 SATMON036 g170772BLASTN 1002 1e−74 90 1508 503 700162040H1 SATMON012 g170772 BLASTN 10031e−74 92 1509 503 700212084H1 SATMON016 g170772 BLASTN 1004 1e−74 891510 503 700027971H1 SATMON003 g170772 BLASTN 982 1e−73 92 1511 503700077321H1 SATMON007 g170772 BLASTN 985 1e−73 89 1512 503 700619884H1SATMON034 g170772 BLASTN 987 1e−73 87 1513 503 700457201H1 SATMON029g170772 BLASTN 988 1e−73 87 1514 503 700082970H1 SATMON011 g170772BLASTN 990 1e−73 89 1515 503 700083350H1 SATMON011 g170772 BLASTN 9711e−72 89 1516 503 700017732H1 SATMON001 g170772 BLASTN 973 1e−72 88 1517503 700094688H1 SATMON008 g170772 BLASTN 975 1e−72 89 1518 503700170985H1 SATMON013 g170772 BLASTN 979 1e−72 89 1519 503 700451847H1SATMON028 g170772 BLASTN 872 1e−71 84 1520 503 700455091H1 SATMON029g170772 BLASTN 931 1e−71 93 1521 503 700106189H1 SATMON010 g170772BLASTN 960 1e−71 88 1522 503 700160075H1 SATMON012 g170772 BLASTN 9681e−71 90 1523 503 700084952H1 SATMON011 g170772 BLASTN 552 1e−70 78 1524503 700348238H1 SATMON023 g170772 BLASTN 946 1e−70 87 1525 503700151310H1 SATMON007 g170772 BLASTN 948 1e−70 90 1526 503 700048962H1SATMON003 g170772 BLASTN 677 1e−69 88 1527 503 700088485H1 SATMON011g170772 BLASTN 936 1e−69 88 1528 503 700093667H1 SATMON008 g170772BLASTN 937 1e−69 88 1529 503 700161491H1 SATMON012 g170772 BLASTN 9281e−68 86 1530 503 700073939H1 SATMON007 g170772 BLASTN 931 1e−68 88 1531503 700027715H1 SATMON003 g170772 BLASTN 472 1e−67 90 1532 503700072843H1 SATMON007 g170772 BLASTN 692 1e−67 87 1533 503 700439356H1SATMON026 g170772 BLASTN 777 1e−67 85 1534 503 700073556H1 SATMON007g170772 BLASTN 910 1e−67 88 1535 503 700154461H1 SATMON007 g170772BLASTN 911 1e−67 87 1536 503 700201909H1 SATMON003 g170772 BLASTN 9121e−67 88 1537 503 700104384H1 SATMON010 g170772 BLASTN 665 1e−66 89 1538503 700152707H1 SATMON007 g170772 BLASTN 898 1e−66 88 1539 503700076625H1 SATMON007 g170772 BLASTN 900 1e−66 85 1540 503 700456945H1SATMON029 g170772 BLASTN 902 1e−66 80 1541 503 700457294H1 SATMON029g170772 BLASTN 446 1e−65 88 1542 503 700152062H1 SATMON007 g170772BLASTN 803 1e−65 90 1543 503 700072428H2 SATMON007 g170772 BLASTN 8901e−65 88 1544 503 700217352H1 SATMON016 g170772 BLASTN 893 1e−65 88 1545503 700072443H2 SATMON007 g170772 BLASTN 894 1e−65 89 1546 503700087436H1 SATMON011 g170772 BLASTN 874 1e−64 88 1547 503 700240615H1SATMON010 g170772 BLASTN 877 1e−64 88 1548 503 700155233H1 SATMON007g170772 BLASTN 883 1e−64 90 1549 503 700241210H1 SATMON010 g170772BLASTN 822 1e−63 87 1550 503 700478070H1 SATMON025 g170772 BLASTN 8631e−63 82 1551 503 700447433H1 SATMON027 g170772 BLASTN 865 1e−63 84 1552503 700215884H1 SATMON016 g170772 BLASTN 865 1e−63 88 1553 503700575250H1 SATMON030 g170772 BLASTN 820 1e−61 84 1554 503 700209283H1SATMON016 g170772 BLASTN 833 1e−60 86 1555 503 700213479H1 SATMON016g170772 BLASTN 834 1e−60 88 1556 503 700241740H1 SATMON010 g170772BLASTN 814 1e−59 88 1557 503 700026012H1 SATMON003 g170772 BLASTN 8151e−59 87 1558 503 700151036H1 SATMON007 g170772 BLASTN 816 1e−59 91 1559503 700207150H1 SATMON017 g170772 BLASTN 818 1e−59 87 1560 503700217383H1 SATMON016 g170772 BLASTN 824 1e−59 88 1561 503 700618168H1SATMON033 g170772 BLASTN 385 1e−58 80 1562 503 700020777H1 SATMON001g170772 BLASTN 806 1e−58 85 1563 503 700224131H1 SATMON011 g170772BLASTN 794 1e−57 88 1564 503 700222876H1 SATMON011 g170772 BLASTN 7941e−57 88 1565 503 700027716H1 SATMON003 g170772 BLASTN 642 1e−56 79 1566503 700073305H1 SATMON007 g170772 BLASTN 787 1e−56 83 1567 503700257376H1 SATMON017 g170772 BLASTN 362 1e−55 87 1568 503 700151752H1SATMON007 g170772 BLASTN 766 1e−55 89 1569 503 700219086H1 SATMON011g170772 BLASTN 769 1e−55 87 1570 503 700218924H1 SATMON011 g170772BLASTN 775 1e−55 87 1571 503 700424527H1 SATMONN01 g170772 BLASTN 7771e−55 83 1572 503 700073183H1 SATMON007 g170772 BLASTN 607 1e−54 85 1573503 700217412H1 SATMON016 g170772 BLASTN 755 1e−54 87 1574 503700208634H1 SATMON016 g170772 BLASTN 642 1e−53 83 1575 503 700202577H1SATMON003 g170772 BLASTN 743 1e−53 86 1576 503 700446645H1 SATMON027g170772 BLASTN 744 1e−53 87 1577 503 700238061H1 SATMON010 g170772BLASTN 745 1e−53 86 1578 503 700617325H1 SATMON033 g170772 BLASTN 7511e−53 91 1579 503 700239901H1 SATMON010 g170772 BLASTN 641 1e−52 86 1580503 700155130H1 SATMON007 g170772 BLASTN 646 1e−52 88 1581 503700083360H1 SATMON011 g170772 BLASTN 739 1e−52 87 1582 503 700236915H1SATMON010 g170772 BLASTN 740 1e−52 87 1583 503 700074780H1 SATMON007g170772 BLASTN 528 1e−51 83 1584 503 700479515H1 SATMON034 g170772BLASTN 718 1e−51 88 1585 503 700215545H1 SATMON016 g170772 BLASTN 7241e−51 90 1586 503 700165354H1 SATMON013 g170772 BLASTN 728 1e−51 86 1587503 700353994H1 SATMON024 g170772 BLASTN 713 1e−50 87 1588 503700153619H1 SATMON007 g170772 BLASTN 694 1e−49 86 1589 503 700074779H1SATMON007 g170772 BLASTN 705 1e−49 77 1590 503 700155530H1 SATMON007g170772 BLASTN 686 1e−48 86 1591 503 700221207H1 SATMON011 g170772BLASTN 692 1e−48 86 1592 503 700264257H1 SATMON017 g170772 BLASTN 6721e−47 87 1593 503 700152216H1 SATMON007 g170772 BLASTN 659 1e−46 88 1594503 700150643H1 SATMON007 g170772 BLASTN 659 1e−46 88 1595 503700156027H1 SATMON007 g170772 BLASTN 655 1e−45 93 1596 503 700260335H1SATMON017 g170772 BLASTN 385 1e−44 79 1597 503 700623795H1 SATMON034g170772 BLASTN 401 1e−44 88 1598 503 700150581H1 SATMON007 g170772BLASTN 640 1e−44 88 1599 503 700151970H1 SATMON007 g170772 BLASTN 6451e−44 87 1600 503 700151780H1 SATMON007 g170772 BLASTN 628 1e−43 88 1601503 700575547H1 SATMON030 g170772 BLASTN 614 1e−42 90 1602 503LIB143-026-Q1-E1-A5 LIB143 g170772 BLASTN 633 1e−42 77 1603 503700347945H1 SATMON023 g170772 BLASTN 586 1e−40 84 1604 503 700074416H1SATMON007 g170772 BLASTN 589 1e−40 86 1605 503 700352990H1 SATMON024g170772 BLASTN 575 1e−39 92 1606 503 700156025H1 SATMON007 g170772BLASTN 568 1e−38 91 1607 503 700432072H1 SATMONN01 g170772 BLASTN 3741e−37 84 1608 503 700354249H1 SATMON024 g170772 BLASTN 528 1e−35 93 1609503 700617977H1 SATMON033 g170772 BLASTN 535 1e−35 87 1610 503700218312H1 SATMON016 g170772 BLASTN 236 1e−33 91 1611 503 700456080H1SATMON029 g170772 BLASTN 513 1e−33 83 1612 503 700349395H1 SATMON023g170772 BLASTN 500 1e−32 91 1613 503 700051637H1 SATMON003 g170772BLASTN 249 1e−31 86 1614 503 700202153H1 SATMON003 g170772 BLASTN 4741e−30 91 1615 503 700256951H1 SATMON017 g170772 BLASTN 454 1e−29 86 1616503 700150542H1 SATMON007 g170772 BLASTN 343 1e−28 76 1617 503700151629H1 SATMON007 g170772 BLASTN 445 1e−28 85 1618 503 700446654H1SATMON027 g170772 BLASTN 437 1e−27 84 1619 503 700155814H1 SATMON007g170772 BLASTN 428 1e−26 88 1620 503 700161019H1 SATMON012 g2588780BLASTN 417 1e−25 92 1621 503 700377236H1 SATMON019 g170772 BLASTN 4021e−24 84 1622 503 LIB3067-036-Q1-K1-A4 LIB3067 g1220121 BLASTN 224 1e−2284 1623 503 700158933H1 SATMON012 g170772 BLASTN 368 1e−21 90 1624 503700155365H1 SATMON007 g170772 BLASTN 347 1e−20 91 1625 503 700405366H1SATMON029 g170772 BLASTN 311 1e−17 88 1626 503 700159812H1 SATMON012g407412 BLASTX 160 1e−15 96 1627 503 700209759H1 SATMON016 g170772BLASTN 239 1e−15 90 1628 503 700154527H1 SATMON007 g170772 BLASTN 2501e−12 92 1629 503 700449637H1 SATMON028 g170772 BLASTN 201 1e−10 78 1630503 700096829H1 SATMON008 g170772 BLASTN 216 1e−9 89 2993 -700661285700661285H1 SOYMON005 g1857024 BLASTX 95 1e−12 100 2994 -700750570700750570H1 SOYMON014 g170772 BLASTN 414 1e−24 81 2995 -700752735700752735H1 SOYMON014 g170772 BLASTN 446 1e−27 78 2996 -700755052700755052H1 SOYMON014 g170772 BLASTN 547 1e−45 74 2997 -700756501700756501H1 SOYMON014 g535583 BLASTN 717 1e−50 84 2998 -700831127700831127H1 SOYMON019 g535583 BLASTN 862 1e−63 83 2999 -700851779700851779H1 SOYMON023 g170772 BLASTN 505 1e−33 77 3000 -700888715700888715H1 SOYMON024 g535583 BLASTN 442 1e−31 91 3001 -700889420700889420H1 SOYMON024 g1220121 BLASTN 893 1e−65 84 3002 -700895218700895218H1 SOYMON024 g407411 BLASTN 816 1e−59 83 3003 -700941379700941379H1 SOYMON024 g170772 BLASTN 424 1e−33 72 3004 -700986855700986855H1 SOYMON009 g170772 BLASTN 701 1e−49 74 3005 -701070484701070484H1 SOYMON034 g407411 BLASTN 362 1e−43 73 3006 -701136279701136279H1 SOYMON038 g170772 BLASTN 651 1e−56 80 3007 -GM16478LIB3054-007-Q1-N1-G3 LIB3054 g2244750 BLASTX 70 1e−27 61 3008 -GM23819LIB3040-019-Q1-E1-C5 LIB3040 g535583 BLASTN 258 1e−10 88 3009 -GM29758LIB3050-016-Q1-E1-B11 LIB3050 g535583 BLASTN 391 1e−42 70 3010 16LIB3030-003-Q1-B1-B11 LIB3030 g3088578 BLASTN 1577 1e−122 87 3011 16LIB3050-023-Q1-K1-H9 LIB3050 g535583 BLASTN 1498 1e−116 86 3012 16LIB3030-003-Q1-B1-F7 LIB3030 g535583 BLASTN 1469 1e−113 86 3013 16LIB3055-005-Q1-N1-C11 LIB3055 g170772 BLASTN 1205 1e−109 84 3014 16LIB3065-011-Q1-N1-A3 LIB3065 g170772 BLASTN 622 1e−107 88 3015 16700652256H1 SOYMON003 g170772 BLASTN 591 1e−90 88 3016 16LIB3065-011-Q1-N1-A4 LIB3065 g170772 BLASTN 1182 1e−89 78 3017 16700653827H1 SOYMON003 g170772 BLASTN 660 1e−87 81 3018 16 701099940H1SOYMON028 g1220121 BLASTN 1154 1e−87 88 3019 16 701003671H1 SOYMON019g170772 BLASTN 1131 1e−85 92 3020 16 700752105H1 SOYMON014 g170772BLASTN 1116 1e−84 91 3021 16 700653057H1 SOYMON003 g170772 BLASTN 9901e−83 88 3022 16 700945531H1 SOYMON024 g535583 BLASTN 1109 1e−83 89 302316 700980013H1 SOYMON009 g535583 BLASTN 1109 1e−83 88 3024 16701127812H1 SOYMON037 g170772 BLASTN 1110 1e−83 91 3025 16LIB3056-014-Q1-N1-F8 LIB3056 g170772 BLASTN 798 1e−81 82 3026 16700653862H1 SOYMON003 g1220121 BLASTN 960 1e−80 88 3027 16 700994148H1SOYMON011 g535583 BLASTN 1069 1e−80 86 3028 16 700984184H1 SOYMON009g170772 BLASTN 756 1e−79 86 3029 16 701123715H1 SOYMON037 g1220121BLASTN 1059 1e−79 87 3030 16 700839038H1 SOYMON020 g170772 BLASTN 10601e−79 93 3031 16 700978445H1 SOYMON009 g170772 BLASTN 1062 1e−79 89 303216 701123035H1 SOYMON037 g170772 BLASTN 829 1e−78 90 3033 16 701041545H1SOYMON029 g170772 BLASTN 1030 1e−77 86 3034 16 700898192H1 SOYMON027g1220121 BLASTN 1031 1e−77 89 3035 16 700985750H1 SOYMON009 g169662BLASTN 1033 1e−77 85 3036 16 700730995H1 SOYMON009 g170772 BLASTN 10401e−77 91 3037 16 700555909H1 SOYMON001 g535583 BLASTN 593 1e−76 84 303816 700746538H1 SOYMON013 g535583 BLASTN 834 1e−76 88 3039 16 700941380H1SOYMON024 g170772 BLASTN 991 1e−76 91 3040 16 701118851H1 SOYMON037g170772 BLASTN 1019 1e−76 89 3041 16 701065379H1 SOYMON034 g535583BLASTN 1022 1e−76 86 3042 16 701209645H1 SOYMON035 g170772 BLASTN 7951e−75 86 3043 16 701055017H1 SOYMON032 g1220121 BLASTN 877 1e−75 86 304416 701015213H1 SOYMON019 g535583 BLASTN 1008 1e−75 87 3045 16700982770H1 SOYMON009 g1220121 BLASTN 1009 1e−75 85 3046 16 701212420H1SOYMON035 g535583 BLASTN 1012 1e−75 86 3047 16 700977916H1 SOYMON009g170772 BLASTN 700 1e−74 89 3048 16 700974401H1 SOYMON005 g1220121BLASTN 857 1e−74 89 3049 16 700645749H1 SOYMON010 g170772 BLASTN 9961e−74 82 3050 16 700646620H1 SOYMON014 g170772 BLASTN 996 1e−74 89 305116 701126103H1 SOYMON037 g170772 BLASTN 1000 1e−74 88 3052 16700978001H1 SOYMON009 g170772 BLASTN 563 1e−73 89 3053 16 700561987H1SOYMON002 g1220121 BLASTN 782 1e−73 87 3054 16 701101526H1 SOYMON028g1220121 BLASTN 810 1e−73 88 3055 16 700562680H1 SOYMON002 g170772BLASTN 985 1e−73 87 3056 16 700560521H1 SOYMON001 g170772 BLASTN 9881e−73 80 3057 16 701055544H1 SOYMON032 g170772 BLASTN 992 1e−73 88 305816 701061418H1 SOYMON033 g170772 BLASTN 993 1e−73 87 3059 16 701049514H1SOYMON032 g1220121 BLASTN 884 1e−72 88 3060 16 700646215H1 SOYMON012g170772 BLASTN 971 1e−72 89 3061 16 701014875H1 SOYMON019 g535583 BLASTN973 1e−72 87 3062 16 700874718H1 SOYMON018 g169662 BLASTN 974 1e−72 863063 16 700897796H1 SOYMON027 g1220121 BLASTN 977 1e−72 87 3064 16700548019H1 SOYMON001 g170772 BLASTN 547 1e−71 87 3065 16 700904875H1SOYMON022 g535583 BLASTN 963 1e−71 88 3066 16 701106315H1 SOYMON036g170772 BLASTN 557 1e−70 90 3067 16 700745023H1 SOYMON013 g407411 BLASTN949 1e−70 85 3068 16 701120348H1 SOYMON037 g170772 BLASTN 956 1e−70 863069 16 701124508H1 SOYMON037 g170772 BLASTN 578 1e−69 84 3070 16700956183H1 SOYMON022 g1220121 BLASTN 625 1e−69 87 3071 16 700894619H1SOYMON024 g535583 BLASTN 828 1e−69 85 3072 16 700892392H1 SOYMON024g535583 BLASTN 936 1e−69 85 3073 16 700730676H1 SOYMON009 g535583 BLASTN939 1e−69 86 3074 16 700728913H1 SOYMON009 g407411 BLASTN 940 1e−69 873075 16 700845737H1 SOYMON021 g1220121 BLASTN 528 1e−68 88 3076 16700990961H1 SOYMON011 g170772 BLASTN 582 1e−68 85 3077 16 700983443H1SOYMON009 g170772 BLASTN 922 1e−68 86 3078 16 701120754H1 SOYMON037g170772 BLASTN 923 1e−68 88 3079 16 700900486H1 SOYMON027 g1220121BLASTN 924 1e−68 83 3080 16 701133574H2 SOYMON038 g170772 BLASTN 9291e−68 88 3081 16 700900924H1 SOYMON027 g170772 BLASTN 931 1e−68 88 308216 701056706H1 SOYMON032 g170772 BLASTN 486 1e−67 89 3083 16 701110051H1SOYMON036 g170772 BLASTN 910 1e−67 88 3084 16 701136325H1 SOYMON038g170772 BLASTN 915 1e−67 87 3085 16 700750809H1 SOYMON014 g170772 BLASTN900 1e−66 88 3086 16 700686607H1 SOYMON008 g170772 BLASTN 901 1e−66 883087 16 700848261H1 SOYMON021 g535583 BLASTN 905 1e−66 87 3088 16700686634H1 SOYMON008 g170772 BLASTN 905 1e−66 88 3089 16 700891285H1SOYMON024 g170772 BLASTN 906 1e−66 88 3090 16 700560291H1 SOYMON001g170772 BLASTN 906 1e−66 88 3091 16 700752975H1 SOYMON014 g170772 BLASTN907 1e−66 89 3092 16 701006013H2 SOYMON019 g170772 BLASTN 908 1e−66 873093 16 700974038H1 SOYMON005 g1220121 BLASTN 631 1e−65 88 3094 16701047024H1 SOYMON032 g170772 BLASTN 707 1e−65 89 3095 16 700900409H1SOYMON027 g170772 BLASTN 737 1e−65 83 3096 16 701137320H1 SOYMON038g170772 BLASTN 769 1e−65 88 3097 16 700978805H1 SOYMON009 g170772 BLASTN886 1e−65 84 3098 16 700726195H1 SOYMON009 g1220121 BLASTN 889 1e−65 873099 16 700661112H1 SOYMON005 g170772 BLASTN 661 1e−64 85 3100 16700989712H1 SOYMON011 g170772 BLASTN 788 1e−64 88 3101 16 700752287H1SOYMON014 g170772 BLASTN 875 1e−64 85 3102 16 700964226H1 SOYMON022g170772 BLASTN 877 1e−64 88 3103 16 700847346H1 SOYMON021 g170772 BLASTN880 1e−64 83 3104 16 700756428H1 SOYMON014 g170772 BLASTN 883 1e−64 883105 16 701049928H1 SOYMON032 g170772 BLASTN 885 1e−64 82 3106 16700848652H1 SOYMON021 g170772 BLASTN 477 1e−63 89 3107 16 700898929H1SOYMON027 g1220121 BLASTN 514 1e−63 87 3108 16 700903523H1 SOYMON022g170772 BLASTN 527 1e−63 82 3109 16 700983745H1 SOYMON009 g1220121BLASTN 863 1e−63 88 3110 16 700890587H1 SOYMON024 g170772 BLASTN 8651e−63 86 3111 16 700969917H1 SOYMON005 g407411 BLASTN 868 1e−63 83 311216 700808487H1 SOYMON024 g170772 BLASTN 870 1e−63 88 3113 16 700749968H1SOYMON013 g170772 BLASTN 871 1e−63 87 3114 16 700751254H1 SOYMON014g170772 BLASTN 575 1e−62 87 3115 16 701014277H1 SOYMON019 g170772 BLASTN739 1e−62 86 3116 16 700853635H1 SOYMON023 g170772 BLASTN 854 1e−62 873117 16 700752357H1 SOYMON014 g170772 BLASTN 856 1e−62 88 3118 16700754523H1 SOYMON014 g170772 BLASTN 856 1e−62 92 3119 16 700982153H1SOYMON009 g170772 BLASTN 400 1e−61 82 3120 16 700958283H1 SOYMON022g170772 BLASTN 839 1e−61 87 3121 16 700980911H1 SOYMON009 g170772 BLASTN842 1e−61 82 3122 16 700788112H1 SOYMON011 g170772 BLASTN 844 1e−61 833123 16 701005927H1 SOYMON019 g170772 BLASTN 849 1e−61 88 3124 16700756443H1 SOYMON014 g170772 BLASTN 849 1e−61 88 3125 16 700658914H1SOYMON004 g1220121 BLASTN 468 1e−60 85 3126 16 701135266H1 SOYMON038g170772 BLASTN 827 1e−60 87 3127 16 700982179H1 SOYMON009 g170772 BLASTN827 1e−60 82 3128 16 700754593H1 SOYMON014 g170772 BLASTN 832 1e−60 813129 16 700831723H1 SOYMON019 g170772 BLASTN 814 1e−59 91 3130 16700986775H1 SOYMON009 g170772 BLASTN 815 1e−59 92 3131 16 700755219H1SOYMON014 g170772 BLASTN 820 1e−59 84 3132 16 701015494H1 SOYMON019g170772 BLASTN 822 1e−59 88 3133 16 701008473H1 SOYMON019 g170772 BLASTN823 1e−59 87 3134 16 700754981H1 SOYMON014 g170772 BLASTN 824 1e−59 883135 16 700870790H1 SOYMON018 g170772 BLASTN 825 1e−59 81 3136 16700833069H1 SOYMON019 g170772 BLASTN 806 1e−58 86 3137 16 700680127H2SOYMON008 g535583 BLASTN 807 1e−58 86 3138 16 701015374H1 SOYMON019g170772 BLASTN 807 1e−58 87 3139 16 700872895H1 SOYMON018 g535583 BLASTN808 1e−58 88 3140 16 701137912H1 SOYMON038 g170772 BLASTN 374 1e−57 833141 16 700984063H1 SOYMON009 g1220121 BLASTN 575 1e−57 79 3142 16700991988H1 SOYMON011 g170772 BLASTN 791 1e−57 82 3143 16 700873915H1SOYMON018 g170772 BLASTN 793 1e−57 88 3144 16 700978721H1 SOYMON009g170772 BLASTN 797 1e−57 78 3145 16 701213679H1 SOYMON035 g170772 BLASTN798 1e−57 88 3146 16 701102954H1 SOYMON028 g170772 BLASTN 799 1e−57 793147 16 700888289H1 SOYMON024 g170772 BLASTN 712 1e−56 91 3148 16700962086H1 SOYMON022 g170772 BLASTN 783 1e−56 88 3149 16 701052227H1SOYMON032 g535583 BLASTN 788 1e−56 86 3150 16 700755177H1 SOYMON014g170772 BLASTN 768 1e−55 88 3151 16 701123136H1 SOYMON037 g170772 BLASTN771 1e−55 82 3152 16 700979067H1 SOYMON009 g170772 BLASTN 776 1e−55 903153 16 700755513H1 SOYMON014 g170772 BLASTN 759 1e−54 92 3154 16700756639H1 SOYMON014 g170772 BLASTN 761 1e−54 81 3155 16 700554077H1SOYMON001 g170772 BLASTN 371 1e−53 86 3156 16 700653194H1 SOYMON003g170772 BLASTN 395 1e−53 86 3157 16 701110348H1 SOYMON036 g170772 BLASTN743 1e−53 81 3158 16 700753973H1 SOYMON014 g170772 BLASTN 743 1e−53 873159 16 701011009H1 SOYMON019 g535583 BLASTN 733 1e−52 81 3160 16700739086H1 SOYMON012 g170772 BLASTN 456 1e−51 87 3161 16 700740140H1SOYMON012 g170772 BLASTN 723 1e−51 89 3162 16 700565040H1 SOYMON002g170772 BLASTN 729 1e−51 74 3163 16 701148186H1 SOYMON031 g535583 BLASTN665 1e−50 85 3164 16 701142734H1 SOYMON038 g535583 BLASTN 670 1e−47 863165 16 700754441H1 SOYMON014 g170772 BLASTN 385 1e−46 93 3166 16701102588H1 SOYMON028 g1220121 BLASTN 662 1e−46 89 3167 16 700900656H1SOYMON027 g535583 BLASTN 635 1e−44 87 3168 16 700974207H1 SOYMON005g535583 BLASTN 641 1e−44 85 3169 16 700982081H1 SOYMON009 g170772 BLASTN494 1e−43 78 3170 16 701130026H1 SOYMON037 g1220121 BLASTN 482 1e−42 863171 16 701009720H1 SOYMON019 g170772 BLASTN 621 1e−42 87 3172 16700962602H1 SOYMON022 g170772 BLASTN 357 1e−40 91 3173 16 700724934H1SOYMON009 g535583 BLASTN 561 1e−40 81 3174 16 700729305H1 SOYMON009g535583 BLASTN 544 1e−39 82 3175 16 701210054H1 SOYMON035 g535583 BLASTN569 1e−38 85 3176 16 700790192H1 SOYMON011 g535583 BLASTN 293 1e−37 833177 16 700984076H1 SOYMON009 g170772 BLASTN 198 1e−35 88 3178 16700726562H1 SOYMON009 g170772 BLASTN 307 1e−34 78 3179 16 701211376H1SOYMON035 g170772 BLASTN 524 1e−34 86 3180 16 700753085H1 SOYMON014g2588780 BLASTN 358 1e−33 76 3181 16 700727993H1 SOYMON009 g535583BLASTN 465 1e−32 84 3182 16 700561072H1 SOYMON001 g170772 BLASTN 4731e−30 83 3183 16 701211464H1 SOYMON035 g170772 BLASTN 464 1e−28 81 318416 701098045H1 SOYMON028 g169660 BLASTN 386 1e−21 78 3185 16 700945233H1SOYMON024 g407412 BLASTX 150 1e−18 87 3186 16 700752655H1 SOYMON014g758247 BLASTX 172 1e−16 94 3187 16 700735356H1 SOYMON010 g758247 BLASTX152 1e−14 100 3188 16 700683995H1 SOYMON008 g758247 BLASTX 106 1e−13 903189 16 700658760H1 SOYMON004 g1857024 BLASTX 123 1e−13 63 3190 16700762885H1 SOYMON015 g1857024 BLASTX 134 1e−13 89 3191 16 700755740H1SOYMON014 g170773 BLASTX 149 1e−13 100 3192 16 700854969H1 SOYMON023g170772 BLASTN 178 1e−12 80 3193 16 701143036H1 SOYMON038 g169661 BLASTX113 1e−8 83 3194 18409 700786561H1 SOYMON011 g535583 BLASTN 992 1e−73 853195 18409 701008057H1 SOYMON019 g535583 BLASTN 831 1e−60 87 3196 18409701037442H1 SOYMON029 g535583 BLASTN 669 1e−46 86 3197 18409 700942865H1SOYMON024 g535583 BLASTN 464 1e−32 86 3198 7322 700651524H1 SOYMON003g170772 BLASTN 466 1e−65 80 3199 7322 700565758H1 SOYMON002 g170772BLASTN 450 1e−63 81

CYSTATHIONINE β-SYNTHASE (EC 4.2.1.22) Seq No. Cluster ID CloneIDLibrary NCBI gi Method Score P-value % Ident 1631 −700025795 700025795H1SATMON003 g1323263 BLASTX 186 1e−25 68 1632 20651 700344783H1 SATMON021g1813975 BLASTX 41 1e−9 53

CYSTATHIONINE γ-LYASE (EC 4.4.1.1) Seq No. Cluster ID CloneID LibraryNCBI gi Method Score P-value % Ident 1633 −700260027 700260027H1SATMON017 g169475 BLASTX 112 1e−10 75 1634 1228 700027629H1 SATMON003g169475 BLASTX 189 1e−19 87 3203 −700750583 700750583H1 SOYMON014g169475 BLASTX 149 1e−13 78 3204 12502 LIB3051-069-Q1-K1-E6 LIB3051g2641242 BLASTX 86 1e−30 38

O-ACETYLHOMOSERINE (THIOL)-LYASE (EC 4.2.99.10) Seq No. Cluster IDCloneID Library NCBI gi Method Score P-value % Ident 3200 12502701135185H1 SOYMON038 g1628606 BLASTX 100 1e−10 48 3201 12502701042913H1 SOYMON029 g2605905 BLASTX 110 1e−9 42 3202 12502 701059330H1SOYMON033 g2605905 BLASTX 110 1e−9 42*Table HeadingsCluster ID

A cluster ID is arbitrarily assigned to all of those clones which belongto the same cluster at a given stringency and a particular clone willbelong to only one cluster at a given stringency. If a cluster containsonly a single clone (a “singleton”), then the cluster ID number will benegative, with an absolute value equal to the clone ID number of itssingle member. The cluster ID entries in the table refer to the clusterwith which the particular clone in each row is associated.

Clone ID

The clone ID number refers to the particular clone in the PhytoSeqdatabase. Each clone ID entry in the table refers to the clone whosesequence is used for (1) the sequence comparison whose scores arepresented and/or (2) assignment to the particular cluster which ispresented. Note that a clone may be included in this table even if itssequence comparison scores fail to meet the minimum standards forsimilarity. In such a case, the clone is included due solely to itsassociation with a particular cluster for which sequences of one or moreother member clones possess the required level of similarity.

Library

The library ID refers to the particular cDNA library from which a givenclone is obtained. Each cDNA library is associated with the particulartissue(s), line(s) and developmental stage(s) from which it is isolated.

NCBI gi

Each sequence in the GenBank public database is arbitrarily assigned aunique NCBI gi (National Center for Biotechnology Information GenBankIdentifier) number. In this table, the NCBI gi number which isassociated (in the same row) with a given clone refers to the particularGenBank sequence which is used in the sequence comparison. This entry isomitted when a clone is included solely due to its association with aparticular cluster.

Method

The entry in the “Method” column of the table refers to the type ofBLAST search that is used for the sequence comparison. “CLUSTER” isentered when the sequence comparison scores for a given clone fail tomeet the minimum values required for significant similarity. In suchcases, the clone is listed in the table solely as a result of itsassociation with a given cluster for which sequences of one or moreother member clones possess the required level of similarity.

Score

Each entry in the “Score” column of the table refers to the BLAST scorethat is generated by sequence comparison of the designated clone withthe designated GenBank sequence using the designated BLAST method. Thisentry is omitted when a clone is included solely due to its associationwith a particular cluster. If the program used to determine the hit isHMMSW then the score refers to HMMSW score.

P-Value

The entries in the P-Value column refer to the probability that suchmatches occur by chance.

% Ident

The entries in the “% Ident” column of the table refer to the percentageof identically matched nucleotides (or residues) that exist along thelength of that portion of the sequences which is aligned by the BLASTcomparison to generate the statistical scores presented. This entry isomitted when a clone is included solely due to its association with aparticular cluster.

1. A substantially purified nucleic acid molecule that encodes a maizeS-adenosyl-methionine decarboxylase fragment wherein said nucleic acidmolecule comprises the nucleic acid sequence of SEQ ID NO:
 662. 2-13.(canceled)
 14. A substantially purified nucleic acid molecule comprisingthe nucleic acid sequence of SEQ ID NO:
 662. 15. (canceled)